Soybean plants having superior agronomic performance and methods for their production

ABSTRACT

This invention provides compositions including favorable alleles of marker loci associated with genetic elements contributing to superior agronomic performance. Also provided are markers for identifying favorable alleles of marker loci associated with genetic elements involved in superior agronomic performance, as well as methods employing the markers.

FIELD OF THE INVENTION

The invention relates to the field of agricultural biotechnology and,more specifically, to molecular marker assisted selection and breedingof soybean, plants.

BACKGROUND OF THE INVENTION

One of the most challenging aspects of plant breeding is to identifyplant varieties that are superior to the currently available varietiesused in commerce. Herein, the term “variety” and “genotype” will be usedinterchangeably since genetic differences are what make each varietyunique and what make one variety superior to another in terms ofcommercial value.

For commodity crops like soybeans and corn, the most universal measureof commercial value is grain productivity per unit area or “yield”.Since a farmer is paid according to the quantity (weight) of grain hedelivers to an elevator, a farmer typically wants to plant a varietythat produces the most grain per acre.

Although yield is arguably the most important trait that a plant breederis concerned with, it is also the least understood genetically. Thereare many different plant traits that control the efficiency ofconverting nutrients and light into grain. Yield is therefore the finalculmination of many different traits that contribute to productivityover the growing season. These would include seedling emergence vigor,photosynthetic ability, disease resistance, ability to mine nutrientsfrom the soil, ability to produce flowers, and ability to shuttlephotosynthate into grain, etc. The genetic bases of these individualtraits that contribute to yield are largely unknown. Each trait thatcontributes to “yield” could be controlled by several or many geneticloci. Therefore, the overall genetic basis for yield is undoubtedly verycomplex. This is just one reason why traditional methods of determiningthe genetic basis of yield have not been very successful. To makeincremental improvements in yield potential, for the most part, plantbreeders are still using the same resource-intensive methods that havebeen in use for the last 80 or more years. Existing varieties arecrossed to produce an array of new genotypes which are then exhaustivelytested over many locations and replications in order to get enough yielddata to differentiate the few consistently superior genotypes. This isone of the most expensive and time-consuming aspects of plant breeding.

During the 1990's, genetic markers linked to genes that contribute toyield emerged as a means to improve efficiency in certain aspects of thebreeding process. These success stories have been limited to traits thatare controlled by relatively few genes that are highly heritable. Inthis case, it is fairly routine to make a reliable association between aDNA sequence and a phenotype that can be confirmed with a greenhouse orfield assay. However, until very recently, it has been extremelydifficult to make reliable associations between specific DNA sequencesand a very complex quantitative trait such as yield.

“Breeding bias,” described in U.S. Pat. No. 5,437,697, which isincorporated herein in its entirety for all purposes, is a unique way todetermine which genetic loci have been affected by extended periods ofrecurrent selection for yield. By comparing the genetic marker profilesof modern high yielding varieties to their most distant ancestors,breeding bias can quickly leverage an entire century of yield data todetermine which specific alleles of which genetic markers have increasedin frequency over time due to selection. Since increased yield has beenthe main criteria for selection, these markers are those most likely tobe associated with yield progress over time. The present inventionprovides genetic markers that are associated with yield performance in avariety of geographic regions, as well as methods for utilizing thesemarkers to efficiently identify soybean lines and sublines withincreased yield.

SUMMARY OF THE INVENTION

The present invention provides representative markers that correspondto, and identify, chromosome segments important for superior agronomicperformance in a variety of geographic regions and growing conditions.The markers described herein are shown to be associated with geneticelements contributing to increased yield in soybean. The markers, andmethods for their use, described herein provide the means for definingand identifying soybean plants with improved yield relative to existingelite lines. Using the markers and methods described herein,identification of residual allelic variation among segregating lines ofsoybeans derived from elite strains can be used to increase theefficiency of the breeding program to develop novel sublines of soybeanwith increased yield relative to existing elite strains.

In a first aspect, the invention provides methods for identifyingsoybean sublines with increased yield relative to existing elite linesof soybeans. The methods of the invention involve detecting at least oneallelic form of a plurality of chromosome segments, each of which a)includes a genetic element contributing to increased yield; and, b)includes or is proximal to a marker locus shown to be associated withincreased yield in soybean.

For example, detecting an allelic form involves identifying at least onefavorable allelic form of a chromosome segment, where the identifiedchromosome segment includes a genetic element contributing to increasedyield and either includes or is proximal to (linked to) a markerselected from the specified set of markers which have been shown to beassociated with yield. The favorable allelic form of a chromosomesegment can be confirmed by identifying a polymorphic marker locusselected from the set, which is segregating in various sublines ofprogeny derived from a progenitor soybean. Yield is assessed in at leasttwo sublines of progeny with different allelic forms of the markerlocus, and a subline of progeny with increased yield relative to theprogenitor soybean is identified.

Exemplary marker loci shown by Breeding Bias analysis to be associatedwith genetic elements that contribute to increased yield in one or moregeographic growing region are included in the following set of markers:Satt684, Satt165, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591,Satt155, Satt385, Satt385, Satt225, Satt236, Satt511, P12390B-1,Satt480, Satt632-TB, Satt233, Satt327, Satt329, Satt508, P10635A-1,Satt409, Satt228, Satt429, Satt426, Satt509, SAT_(—)261, Satt197,Satt519, Satt597, SCT_(—)026, Satt415, Satt583, Satt430, P12198A-1,P8584A-1, Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556,Satt272, Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361,P10639A-1, Satt661-TB, Satt190, SAT_(—)311-DB, Satt338, Satt227,Satt640-TB, Satt422, Satt457, Satt457, Satt557, Satt319, SAT_(—)142-DB,Satt460, P13073A-1, Satt307, SCT_(—)028, Satt433, Satt357, Satt321,Satt267, Satt383, Satt295, Satt203, Satt507, SAT_(—)110, P10620A-1,Satt129, Satt147, Satt216, SAT_(—)351, P10621B-2, Satt701, Satt634,Satt558, Satt266, Satt282, Satt537, Satt506, Satt546, P13072A-1,Satt582, Satt389, Satt461, Satt311, Satt514, Satt464, Satt662, Satt543,Satt186, Satt413, Satt672, P13074A-1, P10624A-1, Satt573, Satt598,Satt204, Satt263, Satt491, Satt602, Satt151, Satt355, Satt452,SAT_(—)273-DB, Satt146, Satt193, Satt569, Satt343, Satt586, Satt040,Satt423, Satt348, Satt595, P10782A-1, P3436A-1, P10598A-1, Satt334,Satt510, Satt510, Satt144, Satt522, Satt522, P9026A-1, P10646A-1,P5219A-1, P7659A-2, Satt570, Satt356, Satt130, Satt115, Satt594,Satt533, Satt303, Satt352, Satt566, Satt199, Satt503, Satt517, Satt191,SAT_(—)117, Satt353, Satt442, Satt279, Satt314, Satt142, Satt181,Satt367, Satt127, SCTT012, Satt270, Satt292, Satt440, P10640A-1,Satt249, SAG1223, SAC1699, SCT_(—)065, Satt596, Satt280, Satt406,Satt380, Satt183, Satt529, Satt431, Satt242, Satt102, Satt441, Satt544,Satt617, Satt240, P10618A-1, Satt475, Satt196, SAT_(—)301, Satt523,Satt418, Satt418, Satt398, Satt497, Satt284, Satt166, Satt448, Satt373,Satt513, P12394A-1, Satt590, Satt567, Satt220; SAG1048, Satt536,Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250, Satt346,Satt336, SAT_(—)330-DB, P13069A-1, P5467A-1, P5467A-2, Satt584,SAT_(—)084, P3050A-2, SAT_(—)275-DB, Satt387, Satt549, Satt660, Satt339,Satt255, Satt257, Satt358, P12396A-1, Satt487, Satt259, Satt259,Satt347, Satt420, Satt576, Satt550, Satt633, Satt262, Satt473, Satt477,Satt581, P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1,P10793A-1, P12391A-1, P13560A-1, P13561A-1, P13561A-1, P2481A-1,S60021-TB, S60048-TB, S60076-TB, S60148-TB, S60149-TB, S60201-TB,S60243-TB, S60326-TB, S60338-TB, S60350-TB, S60361-TB, S60422-TB,S60440-TB, S60446-TB, S60505-TB, S60513-TB, S60519-TB, S60536-TB,S60552-TB, S60585-TB, S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724,SAG1055, Satt040, Satt108, Satt109, Satt111, Satt176, Satt176, Satt219,Satt299, and Satt512. As such, marker loci of the set identifychromosome segments contributing to increased yield. Any number ofadditional marker loci linked to a marker locus selected from the setcan be identified and will function as equivalents in the methods of theinvention.

For example, in some embodiments the favorable allelic form of at leastone chromosome segment, i.e., the allelic form associated with increasedyield, is determined by a) identifying at least one polymorphic markerlocus selected from the set in a plurality of sublines of progeny of aprogenitor soybean, and b) assessing yield in at least two sublimes ofprogeny having different allelic forms of the marker locus.

In other embodiments, the methods of identifying a soybean subline withincreased yield involve detecting at least one allele of a marker locussegregating among progeny of a progenitor soybean. The marker locusincludes at least two alleles, one of which correlates with increasedyield whereas the other allele(s) does not correlate with increasedyield. Increase in yield is measured relative to the mean yield of theprogeny. Optionally, more than one market locus is evaluated. The markerloci are selected from the set of loci consisting of: Satt684, Satt165,Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155, Satt385,Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480, Satt632-TB,Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409, Satt228,Satt429, Satt426, Satt509, SAT_(—)261, Satt197, Satt519, Satt597, SCT026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1, Satt359, P10648A-1,P12105A-1, P10641A-1, Satt168, Satt556, Satt272, Satt020, Satt066,Satt534, P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB, Satt190,SAT_(—)311-DB, Satt338, Satt227, Satt640-TB, Satt422, Satt457, Satt457,Satt557, Satt319, SAT_(—)142-DB, Satt460, P13073A-1, Satt307, SCT 028,Satt433, Satt357, Satt321, Satt267, Satt383, Satt295, Satt203, Satt507,SAT_(—)110, P10620A-1, Satt129, Satt147, Satt216, SAT_(—)351, P10621B-2,Satt701, Satt634, Satt558, Satt266, Satt282, Satt337, Satt506, Satt546,P13072A-1, Satt582, Satt389, Satt461, Satt311, Satt514, Satt464,Satt662, Satt543, Satt186, Satt413, Satt672, P13074A-1, P10624A-1,Satt573, Satt598, Satt204, Satt263, Satt491, Satt602, Satt151, Satt355,Satt452, SAT_(—)273-DB, Satt146, Satt193, Satt569, Satt343, Satt586,Satt040, Satt423, Satt348, Satt595, P10782A-1, P3436A-1, P10598A-1,Satt334, Satt510, Satt510, Satt144, Satt522, Satt522, P9026A-1,P10646A-1, P5219A-1, P7659A-2, Satt570, Satt356, Satt130, Satt115,Satt594, Satt533, Satt303, Satt352, Satt566, Satt199, Satt503, Satt517,Satt191, SAT_(—)117, Satt353, Satt442, Satt279, Satt314, Satt142,Satt181, Satt367, Satt127, SCTT012, Satt270, Satt292, Satt440,P10640A-1, Satt249, SAG1223, SAC1699, SCT 065, Satt596, Satt280,Satt406, Satt380, Satt183, Satt529, Satt431, Satt242, Satt102, Satt441,Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196, SAT_(—)301,Satt523, Satt418, Satt418, Satt398, Satt497, Satt284, Satt166, Satt448,Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt720, SAG1048,Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250,Satt346, Satt336, SAT_(—)330-DB, P13069A-1, P5467A-1, P5467A-2, Satt584,SAT_(—)084, P3050A-2, SAT_(—)275-DB, Satt387, Satt549, Satt660, Satt339,Satt255, Satt257, Satt358, P12396A-1, Satt487, Satt259, Satt259,Satt347, Satt420, Satt576, Satt550, Satt633, Satt262, Satt473, Satt477,Satt581, P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1,P10793A-1, P12391A-1, P13560A-1, P13561A-1, P13561A-1, P2481A-1,S60021-TB, S60048-TB, S60076-TB, S60148-TB, 560149-TB, S60201-TB,S60243-TB, S60326-TB, S60338-TB, S60350-TB, S60361-TB, S60422-TB,S60440-TB, S60446-TB, S60505-TB, S60513-TB, S60519-TB, S60536-TB.S60552-TB, S60585-TB, S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724,SAG1055, Satt040, Satt108, Satt109, Satt111, Satt176, Satt176, Satt219,Satt299, and Satt512.

In some embodiments, the allelic form of between about 10% and about100% of chromosome segments in the set, or in a specified subset of timemarkers relevant in a particular geographic region, as enumerated inTables 3 through 12, are detected. Usually the allelic form of betweenabout 10% and about 90% of the chromosome segments relevant in aparticular geographic region are determined. Commonly, a majority of theallelic forms are determined. In an embodiment, the allelic forms ofessentially all of the chromosome segments are determined.

For example, in a breeding program aimed at developing soybeans withincreased yield in the central growing region (e.g., Iowa) allelic formsof between about 10% and about 100% of the chromosome segments includingor proximal to the markers from the set including Satt642, Satt042,Satt364, Satt454, Satt526, Satt300, Satt591, Satt155, Satt385, Satt511,P12390B-1, Satt632-TB, Satt429, SAT_(—)261, Satt197, P10641A-1, Satt556,Satt534, P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB, Satt190,SAT_(—)311-DB, Satt338, Satt640-TB, Satt557, Satt319, SAT_(—)142-DB,Satt460, Satt433, Satt357, Satt321, Satt295, Satt203, Satt507, Satt129,Satt147, SAT_(—)351, P1062113-2, Satt558, Satt701, Satt634, Satt582,Satt389, Satt464, Satt662, Satt672, Satt573, Satt598, Satt263, Satt602,Satt151, SAT_(—)273-DB, Satt146, Satt193, Satt569, Satt176, Satt343,Satt586, Satt040, Satt595, P10782A-1, Satt334, Satt144, Satt522,Satt570, Satt356, Satt533, Satt199, Satt517, Satt191, SAT 117, Satt279,Satt181, Satt127, Satt970, Satt292, SAG1223, SAC1699, SAT_(—)065,Satt596, Satt406, Satt380, Satt183, Satt529, Satt242, Satt617, Satt240,SAT_(—)301, Satt418, Satt398, Satt497, Satt166, Satt448, Satt373,Satt513, P12394A-1, SAG1048, Satt536, Satt175, Satt677, Satt680,P10615A-1, Satt551, Satt346, Satt336, SAT_(—)330-DB, P13069A-1,P5467A-1, P5467A-2, SAT_(—)084, SAT_(—)275-DB, Satt660, Satt339,P12396A-1, Satt358, Satt487, Satt259, Satt420, Satt576, Satt633,Satt477, Satt581, Satt153, Satt243, P10793A-1, P12391A-1, P12392A-1,P13560A-1, P13561A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB,S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB,S60361-TB, S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB,S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB,S60812-TB, SAC1677, SAC1724, SAG1055, Satt040, Satt111, Satt176, Satt219and Satt299 are determined. In an embodiment, the set of markersincludes Satt684, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591,Satt155, Satt385, Satt632-TB, Satt429, SAT-_(—)261, P10641A-1, Satt556,P10638B-2, Satt399, Satt361, Satt661-TB, Satt190, SAT_(—)311-DB,Satt338, Satt640-TB, Satt557, Satt319, SAT_(—)142-DB, Satt321, Satt203,Satt129, Satt147, SAT_(—)351, P1062113-2, Satt701, Satt634, Satt582,Satt389, Satt464, Satt662, Satt672, Satt573, Satt598, Satt263, Satt151,SAT_(—)273-DB, Satt146, Satt193, Satt569, Satt343, Satt586, Satt040,Satt595, Satt334, Satt144, Satt522, Satt570, Satt356, Satt199, Satt517,Satt191, Sat_(—)117, Satt279, Satt181, Satt127, Satt270, Satt292,SAG1223, SAC1699, Sat_(—)065, Satt596, Satt406, Satt380, Satt183,Satt529, Satt242, Satt617, Satt240, SAT_(—)301, Satt418, Satt398,Satt497, Satt166, Satt448, Satt373, Satt513, P12394A-1, SAG1048,Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, SAT_(—)330-DB,P13069A-1, P5467A-1, P5467A-2, SAT_(—)084, SAT_(—)275-DB, Satt660,Satt339, Satt358, Satt487, Satt487, Satt420, Satt576, Satt633, Satt581,Satt153, Satt243, P10793A-1, P13560A-1, P13561A-1, S60021-TB, S60048-TB,S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt111,Satt219 and Satt299 are determined. In an embodiment, the set of markersincludes Satt684, Satt526, Satt591, Satt385, Satt632-TB, Satt429,SAT-261, P10641A-1, Satt556, P10638B-2, Satt190, SAT_(—)311-DB, Satt338,Satt640-TB, Satt557, SAT_(—)142-DB, Satt321, Satt203, Satt129,SAT_(—)351, Satt701, Satt582, Satt389, Satt464, Satt672, Satt598,Satt343, Satt595, Satt334, Satt144, Satt522, Satt570, Satt356, Satt199,Sat_(—)117, Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223,Sat_(—)065, Satt529, Satt242, Satt617, SAT_(—)301, Satt398, Satt497,Satt166, Satt373, SAG1048, Satt680, P10615A-1, SAT_(—)330-DB, P13069A-1,SAT_(—)275-DB, Satt339, Satt487, Satt420, Satt581 and Satt153.

In other embodiments, the methods of identifying a soybean subline withincreased yield involve detecting at least one allele of a marker locussegregating among progeny of a progenitor soybean. The marker locusincludes at least two alleles, one of which correlates with increasedyield whereas the other allele(s) does not correlate with increasedyield. Increase in yield is measured relative to the mean yield of theprogeny. Optionally, more than one marker locus is evaluated. The markerloci are selected from the set of loci consisting of: Satt684, Satt165,Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155, Satt385,Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480, Satt632-TB,Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409, Satt228,Satt429, Satt426, Satt509, SAT_(—)261, Satt197, Satt519, Satt597,SCT_(—)026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1, Satt359,P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556, Satt272, Satt020,Satt066, Satt534, P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB,Satt190, SAT_(—)311-DB, Satt338, Satt227, Satt640-TB, Satt422, Satt457,Satt457, Satt557, Satt319, SAT_(—)142-DB, Satt460, P13073A-1, Satt307,SCT 028, Satt433, Satt357, Satt321, Satt267, Satt383, Satt295, Satt203,Satt507, SAT_(—)110, P10620A-1, Satt129, Satt147, Satt216, SAT_(—)351,P106211′-2, Satt701, Satt634, Satt558, Satt266, Satt282, Satt537,Satt506, Satt546, P13072A-1, Satt582, Satt389, Satt461, Satt311,Satt514, Satt464, Satt662, Satt543, Satt186, Satt413, Satt672,P13074A-1, P10624A-1, Satt573, Satt598, Satt204, Satt263, Satt491,Satt602, Satt151, Satt355, Satt452, SAT_(—)273-DB, Satt146, Satt193,Satt569, Satt343, Satt586, Satt040, Satt423, Satt348, Satt595,P10782A-1, P3436A-1, P10598A-1, Satt334, Satt510, Satt510, Satt144,Satt522, Satt522, P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570,Satt356, Satt130, Satt115, Satt594, Satt533, Satt303, Satt352, Satt566,Satt199, Satt503, Satt517, Satt191, SAT_(—)117, Satt353, Satt442,Satt279, Satt314, Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270,Satt292, Satt440, P10640A-1, Satt249, SAG1223, SAC1699, SCT 065,Satt596, Satt280, Satt406, Satt380, Satt183, Satt529, Satt431, Satt242,Satt102, Satt441, Satt544, Satt617, Satt240, P10618A-1, Satt475,Satt196, SAT_(—)301, Satt523, Satt418, Satt418, Satt398, Satt497,Satt284, Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt590,Satt567, Satt220, SAG1048, Satt536, Satt175, Satt677, Satt680,P10615A-1, Satt551, Satt250, Satt346, Satt336, SAT_(—)330-DB, P13069A-1,P5467A-1, P5467A-2, Satt584, SAT_(—)084, P3050A-2, SAT_(—)275-DB,Satt387, Satt549, Satt660, Satt339, Satt255, Satt257, Satt358,P12396A-1, Satt487, Satt259, Satt259, Satt347, Satt420, Satt576,Satt550, Satt633, Satt262, Satt473, Satt477, Satt581, P11070A-1,Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1, P10793A-1, P12391A-1,P13560A-1, P13561A-1, P13561A-1, P2481A-1, S60021-TB, S60048-TB,S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60410-TB, S60446-TB,S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt040,Satt108, Satt109, Satt111, Satt176, Satt176, Satt219, Satt299, andSatt512.

In some embodiments, at least one allele of between about 10% and about100% of the markers in the set, or in a specified subset of the markersrelevant in a particular geographic region, as enumerated in Tables 3through 12, are detected. Usually at least one allele of between about10% and about 90% of the marker loci relevant in a particular geographicregion are determined. Commonly, a majority of the marker loci areevaluated. In an embodiment, essentially all of the marker loci areevaluated.

For example, in a breeding program aimed at developing soybeans withincreased yield in the central growing region (e.g., Iowa) at least oneallele of between about 10% and about 100% of the marker loci from theset including Satt642, Satt042, Satt364, Satt454, Satt526, Satt300,Satt591, Satt155, Satt385, Satt511, P12390B-1, Satt632-TB, Satt429,SAT_(—)261, Satt197, P10641A-1, Satt556, Satt534, P1063813-2, Satt399.Satt361, P10639A-1, Satt661-TB, Satt190, SAT_(—)311-DB, Satt338,Satt640-TB, Satt557, Satt319, SAT_(—)142-DB, Satt460, Satt433, Satt357,Satt321, Satt295, Satt203, Satt507, Satt129, Satt147, SAT_(—)351,P10621B-2, Satt558, Satt701, Satt634, Satt582, Satt389, Satt464,Satt662, Satt672, Satt573, Satt598, Satt263, Satt602, Satt151,SAT_(—)273-DB, Satt146, Satt193, Satt569, Satt176, Satt343, Satt586,Satt040, Satt595, P10782A-1, Satt334, Satt144, Satt522, Satt570,Satt356, Satt533, Satt199, Satt517, Satt191, SAT_(—)117, Satt279,Satt181, Satt127, Satt270, Satt292, SAG1223, SAC1699, SAT_(—)065,Satt596, Satt406, Satt380, Satt183, Satt529, Satt242, Satt617, Satt240,SAT_(—)301, Satt418, Satt398, Satt497, Satt166, Satt448, Satt373,Satt513, P12394A-1, SAG1048, Satt536, Satt175, Satt677, Satt680,P10615A-1, Satt551, Satt346, Satt336, SAT_(—)330-DB, P13069A-1,P5467A-1, P5467A-2, SAT″ 084, SAT_(—)275-DB, Satt660, Satt339,P12396A-1, Satt358, Satt487, Satt259, Satt420, Satt576, Satt633,Satt477, Satt581, Satt153, Satt243, P10793A-1, P12391A-1, P12392A-1,P13560A-1, P13561A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB,S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB,S60361-TB, S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB,S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB,S60812-TB, SAC1677, SAC1724, SAG1055, Satt040, Satt111, Satt176, Satt219and Satt299 are determined. In an embodiment, the set of markersincludes Satt684, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591,Satt155, Satt385, Satt632-TB, Satt429, SAT-261, P10641A-1, Satt556,P1063811-2, Satt399, Satt361, Satt661-TB, Satt190, SAT_(—)311-DB,Satt338, Satt640-TB, Satt557, Satt319, SAT_(—)142-DB, Satt321, Satt203,Satt129, Satt147, SAT_(—)351, P1062113-2, Satt701, Satt634, Satt582,Satt389, Satt464, Satt662, Satt672, Satt573, Satt598, Satt263, Satt151,SAT_(—)273-D13, Satt146, Satt193, Satt569, Satt343, Satt586, Satt040,Satt595, Satt334, Satt144, Satt522, Satt570, Satt356, Satt199, Satt517,Satt191, Sat_(—)117, Satt279, Satt181, Satt127, Satt270, Satt292,SAG1223, SAC1699, Sat_(—)065, Satt596, Satt406, Satt380, Satt183,Satt529, Satt242, Satt617, Satt240, SAT_(—)301, Satt418, Satt398,Satt497, Satt166, Satt448, Satt373, Satt513, P12394A-1, SAG1048,Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, SAT_(—)330-DB,P13069A-1, P5467A-1, P5467A-2, SAT_(—)084, SAT_(—)275-DB, Satt660,Satt339, Satt358, Satt487, Satt487, Satt420, Satt576, Satt633, Satt581,Satt153, Satt243, P10793A-1, P13560A-1, P13561A-1, S60021-TB, S60048-TB,S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, 360446-TB,S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt111,Satt219 and Satt299 are determined. In an embodiment, the set of markersincludes Satt684, Satt526, Satt591, Satt385, Satt632-TB, Satt429,SAT-261, P10641A-1, Satt556, P10638B-2, Satt190, SAT_(—)311-DB, Satt338,Satt640-TB, Satt557, SAT_(—)142-D13, Satt321, Satt203, Satt129,SAT_(—)351, Satt701, Satt582, Satt389, Satt464, Satt672, Satt598,Satt343, Satt595, Satt334, Satt144, Satt522, Satt570, Satt356, Satt199,Sat_(—)117, Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223,Sat_(—)065, Satt529, Satt242, Satt617, SAT_(—)301, Satt398, Satt497,Satt166, Satt373, SAG1048, Satt680, P10615A-1, SAT_(—)330-DB, P13069A-1,SAT_(—)275-DB, Satt339, Satt487, Satt420, Satt581 and Satt153.

In some embodiments, the allele correlated with increased yield, andconversely, the allele not correlated with increased yield, aredetermined as follows. At least one polymorphic marker locus having atleast two segregating alleles in a plurality of sublines of progenysoybean plants is selected. The yield is assessed in at least twosublines of progeny with different alleles of the marker. A subline withincreased yield relative to the mean yield of the sublines is thenidentified confirming a correlation between one of the segregatingalleles and increased yield.

ed yield, and conversely, the allele not correlated with increasedyield, are determined as follows. At least one polymorphic marker locushaving at least two segregating alleles in a plurality of sublines ofprogeny soybean plants is selected. The yield is assessed in at leasttwo sublines of progeny with different alleles of the marker. A sublinewith increased yield relative to the mean yield of the sublines is thenidentified confirming a correlation between one of the segregatingalleles and increased yield.

The methods for identifying soybean sublines with increased yieldinvolve detecting at least one allelic form of multiple marker loci.Typically, the number of marker loci is greater than two, and typicallyis between 10% and 100% of the set of marker loci including: Satt684,Satt165, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155,Satt385, Satt3B5, Satt225, Satt236, Satt511, P12390B-1, Satt480,Satt632-TB, Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409,Satt228, Satt429, Satt426, Satt509, SAT_(—)261, Satt197, Satt519,Satt597, SCT 026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1,Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556, Satt272,Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361, P10639A-1,Satt661-TB, Satt190, SAT_(—)311-DB, Satt338, Satt227, Satt640-TB,Satt422, Satt457, Satt457, Satt557, Satt319, SAT_(—)142-DB, Satt460,P13073A-1, Satt307, SCT_(—)028, Satt433, Satt357, Satt321, Satt267,Satt383, Satt295, Satt203, Satt507, SAT_(—)110, P10620A-1, Satt129,Satt147, Satt216, SAT_(—)351, P10621B-2, Satt701, Satt634, Satt558,Satt266, Satt282, Satt537, Satt506, Satt546, P13072A-1, Satt582,Satt389, Satt461, Satt311, Satt514, Satt464, Satt662, Satt543, Satt186,Satt413, Satt672, P13074A-1, P10624A-1, Satt573, Satt598, Satt204,Satt263, Satt491, Satt602, Satt151, Satt355, Satt452, SAT_(—)273-DB,Satt146, Satt193, Satt569, Satt343, Satt586, Satt040, Satt423, Satt348,Satt595, P10782A-1, P3436A-1, P10598A-1, Satt334, Satt510, Satt510,Satt144, Satt522, Satt522, P9026A-1, P10646A-1, P5219A-1, P7659A-2,Satt570, Satt356, Satt130, Satt115, Satt594, Satt533, Satt303, Satt352,Satt566, Satt199, Satt503, Satt517, Satt191, SAT_(—)117, Satt353,Satt442, Satt279, Satt314, Satt142, Satt181, Satt367, Satt127, SCTT012,Satt270, Satt292, Satt440, P10640A-1, Satt249, SAG1223, SAC1699,SCT_(—)065, Satt596, Satt280, Satt406, Satt380, Satt183, Satt529,Satt431, Satt242, Satt102, Satt441, Satt544, Satt617, Satt240,P10618A-1, Satt475, Satt196, SAT_(—)301, Satt523, Satt418, Satt418,Satt398, Satt497, Satt284, Satt166, Satt448, Satt373, Satt513,P12394A-1, Satt590, Satt567, Satt220, SAG1048, Satt536, Satt175,Satt677, Satt680, P10615A-1, Satt551, Satt250, Satt346, Satt336,SAT_(—)330-DB, P13069A-1, P5467A-1, P5467A-2, Satt584, SAT_(—)084,P3050A-2, SAT_(—)275-DB, Satt387, Satt549, Satt660, Satt339, Satt255,Satt257, Satt358, P12396A-1, Satt487, Satt259, Satt259, Satt347,Satt420, Satt576, Satt550, Satt633, Satt262, Satt473, Satt477, Satt581,P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1, P10793A-1,P12391A-1, P13560A-1, P13561A-1, P13561A-1, P2481A-1, S60021-TB,S60048-TB, S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB,S60326-TB, S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB,S60446-TB, S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB,360585-TB, S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055,Satt040, Satt108, Satt109, Satt111, Satt176, Satt176, Satt219, Satt299,and Satt512, or a geographically relevant subset thereof as indicated inTables 3 through 12. Usually, the methods involve detecting betweenabout 10% and about 90% of the markers in the set. Frequently, at least50% of the markers are detected, i.e., a majority of the markers in theset. In some instances essentially all of the markers are detected. Eachof the detected marker loci identifies a chromosome segment shown toinclude a genetic element which contributes to increased yield in atleast one geographic growing region. Thus, by identifying alleles of themarkers associated with increased yield, sublines of soybeans withincreased yield are identified.

The population of progeny soybeans utilized can be obtained by crossinga first progenitor soybean with a second progenitor soybean.Alternatively, the population of progeny can be obtained by selling asingle progenitor soybean. In some embodiments, the sublines of progenyevaluated include random sublines. In some embodiments, the sublinesinclude near isogenic sublines.

Typically, the progenitor soybean is selected from an elite strain ofgermplasm. Such a progenitor can be self-fertilized to generate progeny.More commonly, in the methods of the invention, the progenitor soybeanis crossed to a second soybean selected from a different elite strain ofgermplasm. Alternatively, the progenitor soybean can be crossed to asoybean with an exotic strain of germplasm.

The elite strain of germplasm is typically selected from among thefollowing strains of germplasm: 90A07, 90B11, 90B31, 90B43, 90B72,90B73, 91B01, 91B12, 91B33, 91B52, 91B53, 91B64, 91B91, 91B92, 92B05,92B12, 92B23, 92B38, 92B52, 92B63, 92B74, 92B75, 92B84, 92B95, 92M30,92M31, 92M70, 92M71, 92M72, 92M80, 92M91, 93B01, 93B09, 93B11, 93B15,93B25, 93B26, 93B36, 93B41, 93B45, 93B46, 93B66, 93B67, 93B68, 93B72,93B82, 93B84, 93B85, 93B86, 93B87, 93M10, 93M30, 93M40, 93M50, 93M60,93M80, 93M90, 93M92, 93M93, 94B01, 94B23, 94B24, 94B53, 94B54, 94B73,95B32, 95B33, 95B34, 95B53, 95B95, 95B96, 95B97, 96B21, 96B51, 97B52,97B61, A1395, A2722, A2835, A2943, A3127, A3237, A3242, A3322, A3431,A4009, A4138, A4415, A4595, A4715, A5403, A5560, A5843, A5885, A5979,A5980, A6297, BEDFORD, CM428, CX105, CX232, CX253, CX289, CX394C,CX469C, D00566D362, ESSEX, EX04C00, EX06A00, EX10F01, EX13P01, EX13Q01,EX15N01, EX16N00, EX16P01, EX22Y01, EX22Z01, EX23B03, EX34T03, EX35F03,EX36Y01, EX39E00, EX40T03, EX44V03, FORREST, G3362, HS93-4118,HUTCHESON, JIM, KORADA, MO15733, MO400644-02, MO413735-11-52,MO501577-27-23, MO505469-61-89, MP39009, P1677, P9007, P9008, P9041,P9042, P9061, P9062, P9063, P9071, P9092, P9132, P9141, P9151, P9163,P9182, P9203, P9233, P9244, P9273, P9281, P9305, P9306, P9321, P9341,P9392, P9395, P9481, P9482, P9492, P9521, P9552, P9561, P9584, P9591,P9592, P9594, P9631, P9641, PHARAOH, RA451, R01154R002, S0066, S03W4,S0880, S1550, S1990, S19T9, S20F8, S22C3, S24L2, S25J5, S32Z3, S33N1,S38T8, S3911, S4260, S42H1, S43B5, S5960, S6189, S6262, ST0653, ST1073,ST1090, ST1570, ST1690, ST1970, ST2250, ST2488, ST2660, ST2686, ST2688,ST2788, ST2870, ST3171, ST3380, ST3630, ST3660, ST3870, ST3883, TRACY,TRAILL, X9916, YB03E00, XB03F01, XB07E01, XB10D01, XB15M01, XB19U04,XB20M01, XB22C04, XB22R01, XB23 W03, XB23Y02, XB25E02, XB25L04, XB25×04,XB25W01, XB26L04, XB27P04, XB29A04, XB29D01, XB29K04, XB29L04, XB30E04,XB31C01, XB31R04, XB33B, XB34D04, XB34E01, XB35D, XB35L04, XB35W00,XB38A01, XB41M01, XB42J00, XB42M01, XB48H01, XB54101, XB55J01, XB58P99,XB63D00, XB67A00, YB03G01, YB08D01, YB09F01, YB09G01, YB10E01, YB11D01,YB14H01, YB15K99, YB21F01, YB21G01, YB22S00, YB22V01, YB22W01, YB22X01,YB24Z01, YB25R03, YB25R99, YB25X00, YB25Y01, YB25Z01, YB27L03, YB27S00,YB27X01, YB27Y01, YB28A03, YB28N01, YB29H01, YB29J01, YB29T04, YB30J01,YB30N01, YB30P01, YB31E01, YB32K01, YB33K01, YB34H01, YB34R03, YB34S03,YB35C01, YB36E03, YB36V00, YB38E03, YB38G03, YB39M01, YB39V03, YB40M01,YB40N01, YB41Q01, YB48L01, YB52J00, YB53E00, YB54H00, YB54J00, YB54L00,YB55H00, YB56E00, YB60N01, and YOUNG.

In some embodiments, the methods include electronically transmitting orelectronically storing data representing the determined marker allelesor allelic forms or chromosome segments in a computer readable medium.Accordingly, another aspect of the invention includes computer systemsincluding a data input device for inputting genotyping data, and acomputer readable medium incorporating the genotyping data correspondingto the markers of the invention. Computer readable medium including thegenotyping data are also a feature of the invention.

In some embodiments, the methods further include selecting at least oneplant of the identified soybean subline. The selected plant can be awhole plant, a plant organ, a plant seed, a plant cell, a plant tissueculture, or the like. Optionally, the selected soybean plant, or aprogeny thereof is crossed with a second soybean plant. Typically thesecond soybean plant lacks the determined allele of the marker locus (orallelic form of the chromosome segment). In some embodiments, the secondsoybean plant is from an elite strain of germplasm. In otherembodiments, the second soybean plant is from an exotic strain ofgermplasm.

Soybean plants with increased yield produced according to the methods ofthe invention are also a feature of the invention.

In another aspect, the invention includes sets of markers useful foridentifying soybean plants with increased yield. The marker sets includemarkers selected from the set of markers including Satt684, Satt165,Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155, Satt385,Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480, Satt632-TB,Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409, Satt228,Satt429, Satt426, Satt509, SAT_(—)261, Satt197, Satt519, Satt597, SCT026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1, Satt359, P10648A-1,P12105A-1, P10641A-1, Satt168, Satt556, Satt272, Satt020, Satt066,Satt534, P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB, Satt190,SAT_(—)311-DB, Satt338, Satt227, Satt640-TB, Satt422, Satt457, Satt457,Satt557, Satt319, SAT_(—)142-DB, Satt460, P13073A-1, Satt307,SCT_(—)028, Satt433, Satt357, Satt321, Satt267, Satt383, Satt295,Satt203, Satt507, SAT_(—)110, P10620A-1, Satt129, Satt147, Satt216,SAT_(—)351, P10621B-2, Satt701, Satt634, Satt558, Satt266, Satt282,Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389, Satt461,Satt311, Satt514, Satt464, Satt662, Satt543, Satt186, Satt413, Satt672,P13074A-1, P10624A-1, Satt573, Satt598, Satt204, Satt263, Satt491,Satt602, Satt151, Satt355, Satt452, SAT_(—)273-DB, Satt146, Satt193,Satt569, Satt343, Satt586, Satt040, Satt423, Satt348, Satt595, P10782A4,P3436A-1, P10598A-1, Satt334, Satt510, Satt510, Satt144, Satt522,Satt522, P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570, Satt356,Satt130, Satt115, Satt594, Satt533, Satt303, Satt352, Satt566, Satt199,Satt503, Satt517, Satt191, SAT_(—)117, Satt353, Satt442, Satt279,Satt314, Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270, Satt292,Satt440, P10640A-1, Satt249, SAG1223, SAC1699, SCT 065, Satt596,Satt280, Satt406, Satt380, Satt183, Satt529, Satt431, Satt242, Satt102,Satt441, Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196,SAT_(—)301, Satt523, Satt418, Satt418, Satt398, Satt497, Satt284,Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt590, Satt567,Satt220, SAG1048, Satt536, Satt175, Satt677, Satt680, P10615A-1,Satt551, Satt250, Satt346, Satt336, SAT_(—)330-DB, P13069A-1, P5467A-1,P5467A-2, Satt584, SAT_(—)084, P3050A-2, SAT_(—)275-DB, Satt387,Satt549, Satt660, Satt339, Satt255, Satt257, Satt358, P12396A-1,Satt487, Satt259, Satt259, Satt347, Satt420, Satt576, Satt550, Satt633,Satt262, Satt473, Satt477, Satt581, P11070A-1, Satt153, Satt243,P8230A-1, P10623A-1, P10632A-1, P10793A-1, P12391A-1, P13560A-1,P13561A-1, P13561A-1, P2481A-1, S60021-TB, S60048-TB, S60076-TB,S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB,560350-TB, S60361-TB, S60422-113, S60440-TB, S60446-TB, S60505-TB,S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB,S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt040, Satt108,Satt109, Satt111, Satt176, Satt176, Satt219, Satt299, and Satt512.Typically a subset of markers shown to be relevant in a geographicgrowing region is selected, as indicated in Tables 3 through 12. Forexample, in a breeding program designed to develop strains of soybeanwith increased yield in the Central (e.g., Iowa) region, a set ofmarkers selected from among the following marker set is preferred:Satt642, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155,Satt385, Satt511, P12390B-1, Satt632-TB, Satt429, SAT_(—)261, Satt197,P10641A-1, Satt556, Satt534, P1063813-2, Satt399. Satt361, P10639A-1,Satt661-TB, Satt190, SAT_(—)311-DB, Satt338, Satt640-TB, Satt557,Satt319, SAT_(—)142-DB, Satt460, Satt433, Satt357, Satt321, Satt295,Satt203, Satt507, Satt129, Satt147, SAT_(—)351, P10621B-2, Satt558,Satt701, Satt634, Satt582, Satt389, Satt464, Satt662, Satt672, Satt573,Satt598, Satt263, Satt602, Satt151, SAT_(—)273-DB, Satt146, Satt193,Satt569, Satt176, Satt343, Satt586, Satt040, Satt595, P10782A-1,Satt334, Satt144, Satt522, Satt570, Satt356, Satt533, Satt199, Satt517,Satt191, SAT_(—)117, Satt279, Satt181, Satt127, Satt270, Satt292,SAG1223, SAC1699, SAT_(—)065, Satt596, Satt406, Satt380, Satt183,Satt529, Satt242, Satt617, Satt240, SAT_(—)301, Satt418, Satt398,Satt497, Satt166, Satt448, Satt373, Satt513, P12394A-1, SAG1048,Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt346,Satt336, SAT_(—)330-DB, P13069A-1, P5467A-1, P5467A-2, SAT_(—)084,SAT_(—)275-DB, Satt660, Satt339, P12396A-1, Satt358, Satt487, Satt259,Satt420, Satt576, Satt633, Satt477, Satt581, Satt153, Satt243,P10793A-1, P12391A-1, P12392A-1, P13560A-1, P13561A-1, S60021-TB,S60048-TB, S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB,S60326-TB, 560338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB,S60446-TB, S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB,S60585-TB, S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055,Satt040, Satt111, Satt176, Satt219 and Satt299. In some embodiments themarkers of the set are selected from among: Satt684, Satt042, Satt364,Satt454, Satt526, Satt300, Satt591, Satt155, Satt385, Satt632-TB,Satt429, SAT-_(—)261, P10641A-1, Satt556, P10638B-2, Satt399, Satt361,Satt661-TB, Satt190, SAT_(—)311-DB, Satt338, Satt640-TB, Satt557,Satt319, SAT_(—)142-DB, Satt321, Satt203, Satt129, Satt147, SAT_(—)351,P1062113-2, Satt701, Satt634, Satt582, Satt389, Satt464, Satt662,Satt672, Satt573, Satt598, Satt263, Satt151, SAT_(—)273-DB, Satt146,Satt193, Satt569, Satt343, Satt586, Satt040, Satt595, Satt334, Satt144,Satt522, Satt570, Satt356, Satt1.99, Satt517, Satt191, Sat_(—)117,Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223, SAC1699,Sat_(—)065, Satt596, Satt406, Satt380, Satt183, Satt529, Satt242,Satt617, Satt240, SAT_(—)301, Satt418, Satt398, Satt497, Satt166,Satt448, Satt373, Satt513, P12394A-1, SAG-1048, Satt536, Satt175,Satt677, Satt680, P10615A-1, Satt551, SAT_(—)330-D13, P13069A-1,P5467A-1, P5467A-2, SAT_(—)084, SAT_(—)275-DB, Satt660, Satt339,Satt358, Satt487, Satt487, Satt420, Satt576, Satt633, Satt581, Satt153,Satt243, P10793A-1, P13560A-1, P13561A-1, S60021-TB, S60048-TB,S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt111,Satt219 and Satt299. In certain embodiments the markers are selectedfrom the set including: Satt684, Satt526, Satt591, Satt385, Satt632-TB,Satt429, SAT-_(—)261, P10641A-1, Satt556, P1063813-2, Satt190,SAT_(—)311-DB, Satt338, Satt640-TB, Satt557, SAT_(—)142-DB, Satt321,Satt203, Satt129, SAT_(—)351, Satt701, Satt582, Satt389, Satt464,Satt672, Satt598, Satt343, Satt595, Satt334, Satt144, Satt522, Satt570,Satt356, Satt199, Sat_(—)117, Satt279, Satt181, Satt127, Satt270,Satt292, SAG1223, Sat_(—)065, Satt529, Satt242, Satt617, SAT_(—)301,Satt398, Satt497, Satt166, Satt373, SAG1048, Satt680, P10615A-1,SAT_(—)330-DB, P13069A-1, SAT_(—)275-DB, Satt339, Satt487, Satt420,Satt581 and Satt153.

Typically the set of markers includes between about 10% and about 100%of the markers shown to be relevant in a selected geographic region.Usually, the set includes between about 10% and about 90% of therelevant markers. Frequently, the set includes a majority of therelevant markers. In some embodiments, the set includes essentially allof the markers shown to be relevant in a particular geographic region.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-E. Schematic illustration of genetic map indicating positionsof markers.

FIG. 2. Genetic map indicating position of marker loci associated withyield in Iowa.

DETAILED DESCRIPTION

The present invention provides soybean markers associated with lociimportant for soybean yield. Using methods described in U.S. Pat. No.5,437,697, which is incorporated in its entirety for all purposes, aseries of breeding bias analyses were conducted to identify geneticmarkers that define regions of the soybean genome that are important foryield. Each analysis identified loci that were affected by selection inone of seven different soybean growing regions in North America. Foreach geographic region, an “elite population” of between 38 and 86representative elite lines was chosen by an experienced soybean breeder.Each elite line was evaluated for up to 309 molecular markers todetermine its allelic genotype at each of 309 different genetic locispanning the soybean genome.

In addition to determining the marker genotypes for these elite lines,the most relevant leaf ancestors for each representative elite line weregenotyped with the same 309 genetic markers. A “leaf ancestor” is anancestor for whom the previous parents are unknown, representing anendpoint in the pedigree of each elite line. The breeding bias analysisuses computer simulation and the known pedigree structure between theelite lines and the leaf ancestors to determine the “expected” frequencyof each marker allele within the elite population assuming no bias dueto selection. The expected frequency is merely the average frequencywith which a given allele would be expected in the elite population dueto the rules of random Mendelian segregation. The simulation uses theactual pedigrees of each elite line to determine the path that eachallele must take during the simulated inheritance process. For example,for any diploid biparental cross within a pedigree, the simulationassumes a 50-50 chance that a given parent allele will be passed on to agiven progeny from that cross. By knowing the genotypes of the ancestorsand the pedigree of a given elite line, one can simulate the inheritanceof each allele through a pedigree structure to determine how often thatallele would be expected in the elite line according to a purely randomprocess.

However, plant breeding is not a random process; breeders purposelyselect for characteristics that provide adaptation and high grain yieldin specific geographic regions. By practicing many cycles of geneticrecombination and selection of the best genotypes for a given region,breeders indirectly “bias” the gene pool towards the alleles thatprovide the best grain yield in that locale. Using this logic, anymarker allele that was inherited significantly more frequently thanexpected by random simulation, must reside in a genomic region(chromosome segment) that contributes either directly or indirectly tohigh grain yield.

Following identification of regions of the soybean genome important foryield using the Breeding Bias analysis, specific marker allelesassociated with increased yield can be identified in lines and sublinesof soybeans within a breeding program. Since favorable allelic forms ofchromosome segments are linked to and defined by genetic markers,accurate selection based on genotype can then replace inefficientselection based solely on phenotype. The end result is more efficientprogress towards a genotype with the favorable allelic forms withrespect to yield being fixed within the elite gene pool.

DEFINITIONS

Before describing the present invention in detail, it is to beunderstood that this invention is not limited to particular biologicalsystems, e.g., soybean lines, or reagents, such as particular markers,which can, of course, vary. It is also to be understood that theterminology used herein is for the purpose of describing particularembodiments only, and is not intended to be limiting. As used in thisspecification and the appended claims, the singular forms “a,” “an” and“the” include plural referents unless the content clearly dictatesotherwise. The term “plurality” is used to mean “two or more.” Thus, forexample, reference to “a marker” includes a single marker as well as aplurality of markers, such as two or more markers; and the like.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which the invention pertains. Exemplary methods and materialsare described herein, however, any methods and materials similar orequivalent to those described herein, such as additional or alternativemarkers physically and genetically linked to the markers (and alleles)described herein, can be used in the practice of the present invention.In describing and claiming the present invention, the followingterminology will be used in accordance with the definitions set outbelow.

The term “germplasm” refers to an individual, a group of individuals, ora clone representing a genotype, variety, species or culture, or thegenetic material thereof.

In the context of this disclosure, the term “yield” refers to theproductivity per unit area of a particular plant product of commercialsignificance. For example, yield of soybean is commonly measured inbushels of seed per acre or metric tons of seed per hectare per season.Yield is affected by both genetic and environmental factors.“Agronomics,” “agronomic traits,” and “agronomic performance” refer tothe traits (and underlying genetic elements) of a given plant varietythat contribute to yield over the course of growing season. Individualagronomic traits include emergence vigor, vegetative vigor, stresstolerance, disease resistance, herbicide resistance, branching,flowering, seed set, seed size, seed density, standability,threshability and the like. Yield is, therefore, the final culminationof all agronomic traits.

The term “genetic element” or “gene” refers to a heritable sequence ofDNA, i.e., a genomic sequence, with functional significance. The term“gene” can be used to refer to, e.g., a cDNA and/or an mRNA encoded by agenomic sequence, as well as to that genomic sequence.

“Locus” refers to a specific chromosome location in the genome of aspecies where a specific gene can be found. The term “quantitative traitlocus” or “QTL” refers to a genetic locus with at least two alleles thatdifferentially affect the expression of a phenotypic trait on at leastone genetic background, e.g., in at least one breeding population orsample of progeny.

The term “chromosome segment” designates a contiguous linear span ofgenomic DNA that resides in planta on a single chromosome. The geneticelements or genes located on a single chromosome segment are physicallylinked. In the context of the present invention the genetic elementslocated within a chromosome segment are also genetically linked,typically within a genetic recombination distance of less than or equalto 10 centimorgan (CM). That is, two genetic elements within a singlechromosome segment undergo recombination during meiosis with each otherat a frequency of less than or equal to about 10%.

“Allele” refers to one of two or more different DNA sequences at aspecific locus. In the example of a specific locus where a gene forgrowth habit is located, one allele is a specific DNA sequence that,e.g., codes for determinate growth habit while another allele is adifferent DNA sequence that codes for indeterminate growth habit. A“favorable allele” is the allele at a particular locus that confers, orcontributes to, an agronomically desirable phenotype, e.g., increasedyield. A favorable allele of a marker is an allele associated with afavorable allele at a linked locus which confers or contributes to anagronomically desirable phenotype, e.g., increased yield. A favorableallelic form of a chromosome segment is a chromosome segment including aDNA sequence that contributes to superior agronomic performance at oneor more genetic loci physically located on the chromosome segment.

“Allele frequency” refers to the frequency (proportion or percentage) atwhich an allele is present at a locus within an individual, within aline, or within a population of lines. For example, regarding the allele“A,” diploid individuals of genotype “AA,” “Aa,” or “aa” have allelefrequencies of 1.0, 0.5, or 0.0, respectively. One can estimate theallele frequency within a line by averaging the allele frequencies of asample of individuals from that line. Similarly, one can calculate theallele frequency within a population of lines by averaging the allelefrequencies of lines that make up the population. For a population witha finite number of individuals or lines, an allele frequency can beexpressed as a count of individuals or lines containing the allele.

A “genetic marker” is any qualitatively (discretely) inherited phenotypethat can be used to monitor the segregation of alleles at loci that aregenetically linked to the marker. Genetic markers include visible traitssuch as flower color; enzyme variants such as isozymes and molecularmarkers such as simple sequence repeats (SSRs), single nucleotidepolymorphisms (SNPs), e.g., allele specific hybridization (ASH) markers,restriction fragment length polymorphisms (RFLPs) or randomly amplifiedpolymorphic DNA (RAPDs), etc. Thus, a “marker allele,” alternatively an“allele of a marker locus” is one of a plurality of polymorphicnucleotide sequences at a marker locus.

“Codominant markers” reveal the presence of each allele (two per diploidindividual) at a locus, e.g., SSR, SNP (e.g., ASH), RFLP, AFLP markers.“Dominant markers” reveal the presence of only a single allele perlocus, e.g., RAPD markers. The presence of the dominant marker phenotype(e.g., a band of DNA) is an indication that one allele is present ineither the homozygous or heterozygous condition. The absence of thedominant marker phenotype (e.g., absence of a DNA band) is merelyevidence that some other, undefined, allele is present. In the case ofpopulations where individuals are predominantly homozygous and loci arepredominantly dimorphic, dominant and codominant markers are equallyvaluable. As individuals within populations become more heterozygous andmulti-allelic, codominant markers become more informative of genotypethan dominant markers.

A “set” of markers refers to a collection or group of markers, or thedata derived therefrom; used for a common purpose, identifying soybeanplants with increased yield. Frequently, the data is stored in anelectronic medium. While each of the members of the set has been shownto possess utility with respect to the specified purpose: individualmarkers selected from the set as well as subsets including some, but notall of the markers, are also effective in achieving the specifiedpurpose.

A “genetic map” is a description of the genetic linkage relationshipsamong loci on one or more chromosomes (or linkage groups) within a givenspecies, generally depicted in a diagrammatic or tabular form. “Mapping”is the process of defining the linkage relationships of loci through theuse of genetic markers, populations segregating for the markers, andstandard genetic principles of recombination frequency. A “map location”is an assigned location on a genetic map relative to linked geneticmarkers where a specified marker can be found within a given species.

A “genotype” is the genetic constitution of an individual (or group ofindividuals) at one or more genetic loci. Genotype is defined by theallele(s) of one or more known loci that the individual has inheritedfrom its parents. A “haplotype” is the genotype of an individual at aplurality of genetic loci. Typically, the genetic loci described by ahaplotype are physically and genetically linked, i.e., on the samechromosome segment.

An individual is “homozygous” if the individual has only one type ofallele at a given locus (e.g., a diploid individual with two copies ofthe same allele at a locus). An individual is “Heterozygous” if morethan one allele type is present at a given locus (e.g., a diploidindividual with one copy each of two different alleles). The term“homogeneity” indicates that members of a group have the same genotypeat one or more specific loci. In contrast, the term “heterogeneity” isused to indicate that individuals within the group differ in genotype atone or more specific loci.

A “line” or “strain” is a group of individuals of identical parentagethat are generally inbred to some degree and are generally homozygousand homogeneous at most loci.

An “elite line” or “elite strain” is a genetically superior line thathas resulted from many cycles of breeding and selection for superioragronomic performance. Numerous elite lines are available and known tothose of skill in the art of soybean breeding. An “elite population” isan assortment of elite lines that can be used to represent the state ofthe art in terms of agronomically superior genotypes of a given cropspecies, such as soybean. Similarly, an “elite germplasm” or elitestrain of germplasm is a genetically superior germplasm, typicallyderived from and/or capable of giving rise to a plant with superioragronomic performance, such as an existing or newly developed elite lineof soybean.

In contrast, an “exotic germplasm” is a germplasm derived from a soybeannot belonging to an available elite soybean line or strain of germplasm.In the context of a cross between two soybean plants or strains ofgermplasm, an exotic germplasm is unrelated by descent to the elitegermplasm with which it is crossed. Most commonly, the exotic germplasmis not derived from any known elite line of soybean, but rather isselected to introduce novel genetic elements (novel alleles) into abreeding program.

An “ancestral line” is a parent used as a source of genes for thedevelopment of elite lines. An “ancestral population” is a group ofancestors that have contributed the bulk of the genetic variation thatwas used to develop elite lines. “Descendants” are the progeny ofancestors, and may be separated from their ancestors by many generationsof breeding. For example, elite lines are the descendants of theirancestors, A “pedigree structure” defines the relationship between adescendant and each ancestor that gave rise to that descendant. Apedigree structure can span one or more generations, describingrelationships between the descendant and it's parents, grand parents,great-grand parents, etc.

The term “sublime” refers to a an inbred subset of descendents thatgenetically distinct from other similarly inbred subsets descended fromthe same progenitor. Traditionally, a “sublime” has been derived byinbreeding the seed from an individual soybean plant selected and at theF3 to F5 generation until the residual segregating loci are “fixed” orhomozygous across most or all loci. Commercial soybean varieties (orlines) are typically produced by aggregating (“bulking”) theself-pollinated progeny of a single F3 to F5 plant from a controlledcross between 2 genetically different parents. While the varietytypically appears uniform, the self-pollinating variety derived from theselected plant eventually (e.g., F8) becomes a mixture of homozygousplants that can vary in genotype at any locus that was heterozygous inthe originally selected F3 to F5 plant. In the context of the invention,marker-based sublines, that differ from each other based on qualitativepolymorphism at the DNA level at one or more specific marker loci, arederived by genotyping a sample of seed derived from individualself-pollinated progeny derived from a selected F3-F5 plant. The seedsample can be genotyped directly as seed, or as plant tissue grown fromsuch a seed sample. Optionally, seed sharing a common genotype at thespecified locus (or loci) are bulked providing a subline that isgenetically homogenous at identified loci important for increased yield.

The term “near-isogenic” lines refers to lines that are geneticallysimilar to each other except at one or a small number of genetic loci(e.g., at 1, 2, or about 5 to about 10 specified genetic loci). Thesecan be created as described for marker-based sublines or based ondifferences for any qualitative trait that can serve as an effectivegenetic marker. Percent similarity between near-isogenic lines is afunction of the similarity of the parents of the original cross and thegeneration at which self-pollination is performed. On average, therelatedness between members of a given inbred line increases 50% witheach cycle of inbreeding, due to a 50% increase in homozygosity at eachcycle of inbreeding. Percent similarity can be more accuratelydetermined with genetic markers that span the genome. In some cases,near-isogenic lines differ from each other at one defined genetic locus.

“Transgressive segregation” is an inheritance pattern that results in aphenotype (e.g., agronomic performance) of an individual that is moreextreme than either parent. For example, with respect to agronomicperformance, transgressive segregation results in a progeny with yieldthat is greater than a best parent or less than a worst parent.Desirable transgressive segregation is the case where the progeny arebetter than either parent. Transgressive segregation can also bemeasured in terms of the number of favorable alleles that an individualinherits in relation to the number of favorable alleles of each of itsparents. A “target segregant” is a progeny from a specific cross thatincludes only favorable alleles at each defined locus segregating in thecross. The target segregant therefore, has the best possible genotypethat can result from a cross between parents that differ in genotype atknown loci. “Target genotype” refers to an individual containing thefavorable allelic forms at all chromosome segments or loci known toaffect a particular trait or phenotype, such as agronomic performance.With respect to agronomic performance, the target genotype is that ofthe target segregant from a cross between parents that complement interms of favorable alleles at all defined loci affecting agronomicperformance.

A “survey” or “genetic survey” or “genetic marker survey” is the processof determining and recording the genotype of individuals or lines (e.g.,ancestral and elite lines), at any number of defined loci, with the useof genetic markers.

The term “associated with” or “associated,” when referring to a nucleicacid (e.g., a genetic marker) and a phenotype in the context of thepresent invention, refers to a nucleic acid and a phenotypic trait thatare in linkage phase disequilibrium. The term “linkage phasedisequilibrium” or “linkage disequilibrium” refers to a non-randomsegregation of genetic loci. This implies that such loci are insufficient physical proximity along a length of a chromosome that theytend to segregate together with greater than random frequency.

The term “genetically linked” refers to genetic loci (including geneticmarker loci) that are physically close enough to each other on the samechromosome such that they have a recombination frequency of less than0.5. When referring to the relationship between two genetic elements,such as a genetic element contributing to yield and a proximal marker,“Coupling” phase linkage indicates the state where the “favorable”allele at the yield locus is physically associated on the samechromosome strand as the “favorable” allele of the respective linkedmarker locus. In coupling phase, both favorable alleles are inheritedtogether by progeny that inherit that chromosome strand. In “repulsion”phase linkage, the “favorable” allele at the yield locus is physicallylinked with an “unfavorable” allele at the proximal marker locus, andthe two “favorable” alleles are not inherited together (i.e., the twoloci are “out of phase” with each other).

The term “physically linked” is used to indicate that two genetic loci,e.g., two marker loci, a marker locus and a locus contributing tovariation in a phenotype, are physically present on the same chromosome.Typically, the two loci are located in close proximity, such thatrecombination between homologous chromosome pairs does not occur betweenthe two loci with high frequency. That is, recombination between twophysically linked loci typically occurs with a frequency of less thanabout 10%, favorably with a frequency of less than 5%, more favorablywith a frequency of 2% or less or a frequency of 1% or less. Thus, twoloci that are localized to the same chromosome, and at such a distancethat recombination between the two loci occurs at a frequency of lessthan 10% are said to be “proximal to” each other.

“Marker Assisted Selection” or “MAS” refers to the practice of selectingfor desired phenotypes among members of a breeding population usinggenetic markers.

The phrase “hybrid plants” refers to plants which result from a crossbetween genetically different individuals.

The term “crossed” or “cross” in the context of this invention means thefusion of gametes, e.g., via pollination to produce progeny (i.e.,cells, seeds, or plants) in the case of plants. The term encompassesboth sexual crosses (the pollination of one plant by another) and, inthe case of plants, selfing (self-pollination, i.e., when the pollen andovule are from the same plant).

“Random mating” is the mating of individuals within a population in away that insures the equal probability of any two individuals matingregardless of genotype. “Non-random mating” is any deviation from randommating in which specific crosses between individuals occur with greaterfrequency than others.

The term “introgression” refers to the transmission of a desired alleleof a genetic locus from one genetic background to another. For example,introgression of a desired allele at a specified locus can betransmitted to at least one progeny via a sexual cross between twoparents of the same species, where at least one of the parents has thedesired allele in its genome. Alternatively, for example, transmissionof an allele can occur by recombination between two donor genomes, e.g.,in a fused protoplast, where at least one of the donor protoplasts hasthe desired allele in its genome. The desired allele can be, e.g., aselected allele of a marker or QTL or a transgene.

The terms “nucleic acid,” “polynucleotide,” “polynucleotide sequence”and “nucleic acid sequence” refer to single-stranded or double-strandeddeoxyribonucleotide or ribonucleotide polymers, or chimeras thereof. Asused herein, the term can additionally or alternatively include analogsof naturally occurring nucleotides having the essential nature ofnatural nucleotides in that they hybridize to single-stranded nucleicacids in a manner similar to naturally occurring nucleotides (e.g.,peptide nucleic acids). Unless otherwise indicated, a particular nucleicacid sequence of this invention optionally encompasses complementarysequences, in addition to the sequence explicitly indicated.

The term “homologous” refers to nucleic acid sequences that are derivedfrom a common ancestral gene through natural or artificial processes(e_g, are members of the same gene family), and thus, typically, sharesequence similarity. Typically, homologous nucleic acids have sufficientsequence identity that one of the sequences or its complement is able toselectively hybridize to the other under selective hybridizationconditions. The term “selectively hybridizes” includes reference tohybridization, under stringent hybridization conditions, of a nucleicacid sequence to a specified nucleic acid target sequence to adetectably greater degree (e.g., at least 2-fold over background) thanits hybridization to non-target nucleic acid sequences and to thesubstantial exclusion of non-target nucleic acids. Selectivelyhybridizing sequences have about at least 80% sequence identity,preferably at least 90% sequence identity, and most preferably 95%, 97%,99%, or 100% sequence identity with each other. A nucleic acid thatexhibits at least some degree of homology to a reference nucleic acidcan be unique or identical to the reference nucleic acid or itscomplementary sequence.

The term “isolated” refers to material, such as a nucleic acid or aprotein, which is partially or substantially free from components thatnormally accompany or interact with it in its naturally occurringenvironment. The isolated material optionally comprises material notfound with the material in its natural environment, e.g., a cell. Inaddition, if the material is in its natural environment, such as a cell,the material has been placed at a location in the cell (e.g., genome orsubcellular organelle) not native to a material found in thatenvironment. For example, a naturally occurring nucleic acid (e.g., apromoter) is considered to be isolated if it is introduced bynon-naturally occurring means to a locus of the genome not native tothat nucleic acid. Nucleic acids which are “isolated” as defined herein,are also referred to as “heterologous” nucleic acids.

The term “recombinant” when referring to a molecular species, such as anucleic acid or protein, indicates that the material (e.g., a nucleicacid or protein) has been synthetically (non-naturally) altered by humanintervention. The alteration to yield the synthetic material can beperformed on the material within or removed from its natural environmentor state. For example, a naturally occurring nucleic acid is considereda recombinant nucleic acid if it is altered, or if it is transcribedfrom DNA which has been altered, by means of human intervention, e.g.,performed on the cell from which it originates. When the term“recombinant” is used in the classical genetic sense, it refers to anindividual with one of the many possible rearrangments of genes due tonatural sexual recombination. For example, “recombinant inbred lines” or“RILs” are merely the variety of inbred progeny from specific crosses ofdivergent parents. The manner in which the term recombinant is employedwill be self evident from the context of its use.

The term “introduced” when referring to a heterologous or isolatednucleic acid refers to the incorporation of a nucleic acid into aeukaryotic or prokaryotic cell where the nucleic acid can beincorporated into the genome of the cell (e.g., chromosome, plasmid,plastid or mitochondrial DNA), converted into an autonomous replicon, ortransiently expressed (e.g., transfected mRNA). The term includes suchnucleic acid introduction means as “transfection,” “transformation” and“transduction.”

The term “host cell” means a cell that contains a heterologous nucleicacid, such as a vector, and supports the replication and/or expressionof the nucleic acid. Host cells may be prokaryotic cells such as E.coli, or eukaryotic cells such as yeast, insect, amphibian, or mammaliancells. In the context of the present invention, a eukaryotic host cellis most commonly a soybean cell.

The term “transgenic” plant (or animal) refers to a plant (or animal)which comprises within its genome a heterologous polynucleotide.Generally, the heterologous polynucleotide is stably integrated withinthe genome such that the polynucleotide is passed on to successivegenerations. The heterologous polynucleotide may be integrated into thegenome alone or as part of a recombinant expression cassette.“Transgenic” is used herein to refer to any cell, cell line, tissue,part or organisms, the genotype of which has been altered by thepresence of heterologous nucleic acid including those transgenicorganisms or cells initially so altered, as well as those created bycrosses or asexual propagation from the initial transgenic organism orcell. The term “transgenic” as used herein does not encompass thealteration of the genome (chromosomal or extra-chromosomal) byconventional breeding methods (i.e., crosses) or by naturally occurringevents such as random cross-fertilization, viral infection withnon-recombinant nucleic acids, bacterial transformation withnon-recombinant nucleic acids, transposition with non-recombinantnucleic acids, or spontaneous mutation. Examples of processes by which atransgenic organism can be produced are described below, and includeelectroporation, microinjection, Agraobacterium-mediated transformation,biolistic methods, in planta techniques, and the like.

The term “plant” includes any of: whole plants, plant organs (e.g.,leaves, stems, roots, etc.), tissues, seeds, plant cells, and/or progenyof the same. Similarly, “plant cell,” as used herein includes, withoutlimitation, seeds, suspension cultures, embryos, meristematic regions,callus tissue, leaves, roots, shoots, gametophytes, sporophytes, pollen,and microspores. In addition, the term “plant” encompasses in silicorepresentations of part or all of a plant's genetic constitution.

Introduction

The present invention addresses the need in the field of agriculture tomore efficiently develop soybean plants having superior agronomicperformance. Superior agronomic performance, as measured by increasedyields of soybeans, is a function of such diverse factors as seedlingemergence vigor, resistance to environmental stress, pest resistance,insect resistance, disease resistance, ability to mine nutrients fromthe soil, ability to produce flowers, photosynthetic ability, andability to shuttle photosynthate into grain, etc. As disclosed herein,genetic loci associated with superior agronomic performance have beenidentified by analysis of a plurality of soybean breeding programs (someof which extend back more than 70 years) in a variety of geographiczones in the United States and Canada.

Soybean germplasm having superior agronomic performance (relative totheir parents and sibs developed in these same breeding programs) show astatistically significant retention of particular allelic forms(alternatively, “alleles”) encompassed within the chromosome segments ofthe present invention. This highly significant statistical associationdemonstrates that loci within these chromosome segments are linked(genetically and physically) to genetic elements associated with thevarious traits involved in determining soybean yield in the context ofmodern agricultural practices. Collectively, favorable attributescorresponding to such traits are descriptively referred to as “superioragronomic performance,” and contribute to increased yield. Consequently,molecular markers localized within these chromosome segments can be usedto define (and identify) soybean plants with superior agronomicperformance, and in marker assisted selection (“MAS”) and markerassisted breeding strategies (alternatively referred to as “molecularbreeding”) to create soybean plants with superior agronomic performance.Furthermore, chromosome segments encompassing the genetic elementsassociated with the phenotype of superior agronomic performance can beisolated and transformed into soybean or other monocot or dicot plantspecies.

The methodology used to identify these chromosome segments, referred toas “Breeding Bias” is disclosed in U.S. Pat. No. 5,437,697 to Sebastianet al., incorporated herein in its entirety for all purposes.

Briefly, loci (e.g., markers and chromosome segments encompassinggenetic elements contributing to superior agronomic performance) thathave been affected by selection for agronomic performance, areidentified by comparing the genotype of modern elite lines with that oftheir ancestors. Because domesticated soybeans are known to have afairly narrow gene pool and have been selected to fairly stringentstandards, a relatively small number of elite lines is adequate toidentify relevant chromosome regions associated with superior agronomicperformance. The identity of elite lines used for each geographicalregion is indicated in Table 1.

Based on the selection of elite lines, the relevant ancestralpopulation, e.g., ancestors that were used most frequently by breedersto develop the elite population is identified. A pedigree tracing therelationship between each elite line and its earliest known ancestors isobtained, and the genetic contribution of each ancestor is convertedinto a proportional or percentage representation, assuming that onaverage 50% of each parent's genome is passed on to each progeny as aresult of a two-way cross with another parent. By tracing the pedigreeback until no more branch points are found, the earliest known ancestorscan be identified and their contribution to each elite line calculated.Calculations are performed, generally in a computerized format, for eachof the elite lines included in the genetic survey.

Once the appropriate elite and ancestral lines have been chosen, thegenotype of each line is determined through the use of genetic markers.Genetic markers include any qualitative phenotype that can be used as adirect measure of genotype a specific locus. Markers include visualtraits such as flower color, enzyme variants such as isozymes, andmolecular markers, which can be detected by a variety of means such assimple sequence repeats (SSR), single nucleotide polymorphism (SNP),allele specific hybridization (ASH), restriction fragment lengthpolymorphism (RFLP), amplified variable sequences, single strandconformation polymorphism (SSCP), amplified fragment length polymorphism(RFLP) and randomly amplified polymorphic DNA (RAPD).

Regardless of which genetic markers are used to monitor genotype, theend result is a marker genotype for each of the elite and ancestrallines. The genotype of each line is merely an indication of which allelethe line possesses at any number of loci defined by the genetic markers.

Once one has determined the genotypes of ancestral and elite lines,statistical analyses are used to determine whether selection forsuperior agronomic performance has favored particular alleles at certainloci. The first statistic to calculate is the probability of findingeach allele within the elite population with the assumption thatselection had no effect on allele frequency. This expected allelefrequency within the elite population serves as a basis for comparisonto the observed allele frequency.

Expected allele frequency within the elite population is a function ofthe genotype of each ancestor and the pedigrees of elite linesrepresenting the elite population. In a random mating population, theallele frequency among descendants should be similar to allele frequencyamong ancestors unless breeding and selection has favored particularalleles. However, since breeding of many crops (including soybeans) isnot done through random mating, one can use the pedigree of eachdescendant (e.g., elite line) to calculate the probability of inheritinga given allele from its ancestors. Within non-random mating populations,expected allele frequency can be obtained by averaging the individualprobabilities of inheriting an allele over any number of descendants(that may differ greatly in pedigree).

By comparing the observed frequency of a given allele in the elitepopulation to the average probability of inheriting that allele (i.e.,comparing observed count to expected count), one can determine whichloci have been affected by historical selection for agronomic traits.Favorable alleles are identified as the ones that have been inheritedmore frequently than expected (i.e., have been favored by selection).Unfavorable alleles are those inherited less frequently than expected(i.e., selected against). A statistical test is then used, e.g., asdescribed in detail in U.S. Pat. No. 5,437,697, to determine thesignificance of a difference between observed and expected allelefrequency.

According to these methods, loci and alleles with significant deviationsfrom expected allele frequency correlating with superior agronomicperformance in different growing environments have been identified.These loci can be used to define and identify, as well as select forsoybeans with superior agronomic performance. For example, the markersdisclosed herein can be used to 1) identify soybean germplasm withsuperior and/or improved agronomic performance, e.g., relative topresently existing or parental soybean lines; 2) identify parents thatwill produce superior transgressive segregants; 3) select superior linesfrom crosses that are segregating at loci (e.g., QTL loci) whichcontribute to superior agronomic performance; 4) select parents thatwill produce the best hybrids; 5) purify heterogeneous lines, i.e., byselecting only those individuals that include the favorable allele(s) atloci that are still segregating within the line; 6) select for andmaintain desirable heterogeneity; 7) maintain favorable alleles atmultiple loci that have been assembled by many years of selection whileincorporating exotic alleles from new germplasm at other loci; 8) totest the effects of exotic allele substitution at loci that have provenimportant for domestication, e.g., in the event that an exotic alleleprovides better agronomic performance at the loci identified by breedingbias; and, 9) in any process where it is important to prioritize whichloci are more important for agronomic fitness than random loci withinthe genome.

Markers and Alleles Correlating with Superior Agronomic Performance inSoybeans

In one aspect, the present invention provides marker loci correlatedwith superior agronomic performance in soybean. Each of the identifiedmarkers is expected to be in close physical and genetic proximity (i.e.,physically and genetically linked) to a genetic element, e.g., aquantitative trait locus or QTL, that contributes to superior agronomicperformance. If a particular marker were not in proximity to animportant QTL, the extended period of recurrent selection (70+ years forsoybean) would have provided many opportunities for crossing overbetween the marker and QTL. This would result in disassociation betweenthe marker allele and the QTL allele resulting in “linkage equilibrium”and statistically non-significant estimates of correlation between themarker and yield, i.e., LOD or probability scores. Breeding bias doesnot detect poorly-linked markers, that is, markers that are distant fromimportant loci, even if both the marker and the genetic element (e.g.,QTL) contributing to yield are found on the same chromosome. Inaddition, since thousands of different environments have been sampled totest performance during each cycle of selection, alleles that provideadaptation to a wide range of environments will be most readilyidentified. Alleles that are only favorable under rare environmentalconditions will not consistently increase in frequency due to selectionin different environments and will, therefore, not be detected bybreeding bias analysis. Furthermore, the narrow gene pool and highrelatedness among elite soybean germplasm shared by both public andprivate institutions (Delanney et al, 1983, Crop Science 23:944-949) actto homogenize the gene pool and make marker-QTL associations morereliable. Taken together, these features in combination with the largebody of data analyzed by breeding bias, ensure that QTL with asubstantial contribution to yield are located in close proximity to themarkers disclosed herein.

Accordingly, each of the disclosed markers defines a chromosome segmentassociated with a genetic element, e.g., a QTL contributing to superioragronomic performance. One of skill in the art will recognize that foreach of the chromosome segments encompassing QTL related to yield,identification of and/or selection for the QTL is optimized by using agenetic marker that is as close as possible to the actual QTL locus thatis responsible for the phenotype in question. Thus, a “perfect” markerwould be one that is localized within the genetic element or QTL itself,and corresponds to the DNA polymorphism that is responsible for thesuperior phenotype. That is, the marker polymorphism is the mutationunderlying the improvement in yield. However, since most of the geneticmarkers available for soybean consist of RFLPs, SSRs, RAPDs and SNPs ofunknown function, it would be highly unlikely that a given marker fromthose currently available was already “perfect.” Nevertheless, themarkers described herein constitute a set of tools for detectingimportant chromosome segments comprising QTL associated with yield.These markers are sufficiently close to their respective linked QTL todetect a statistically significant shift in allele frequency (due toselection) over 70 years of recurrent selection for yield, thus, areuseful for the purposes of identifying desirable germplasm and for MAS.In addition, these markers are useful for identifying additionalmarkers, e.g., including a “perfect” marker within the QTL gene ofinterest.

Tables 3 through 12 disclose exemplary molecular, which define thechromosome segments that are embodiments of the present invention. Eachof Tables 3 through 12 provides marker loci and favorable alleles,identified by Breeding Bias, relevant in a specified growing environmentdistinguished by geographic region.

The exemplary markers provided in Tables 3 through 12 identifychromosome segments including genetic elements (genes) important foryield in soybean. The chromosome segments of the present invention arecontiguous lengths of chromosome delimited by a specified crossoverfrequency or map distance (centimorgans or CM) from a molecular markerof the present invention. The chromosome segments of the presentinvention are delimited by a crossover frequency of up to about 10%,i.e., 10 CM, from a marker locus known to be associated with superioragronomic performance, e.g., Satt165; Satt042; Satt364; Satt454;Satt526; Satt300; Satt591; Satt155; Satt385; Satt511; P12390B-1;Satt327; Satt329; Satt508; P10635A-1; Satt409; Satt228; Satt429;Satt509; Satt197; SCT 026; Satt415; Satt583; Satt430; P12198A-1;P8584A-1; Satt359; P10648A-1; P10641A-1; Satt168; Satt556; Satt272;Satt020; Satt534; P10638B-2; Satt399; Satt361; P10639A-1; Satt190;Satt338; Satt227; Satt457; Satt557; Satt319; Satt460; Satt307; SCT 028;Satt433; Satt357; Satt321; Satt267; Satt295; Satt203; Satt507;SAT_(—)110; P10620A-1; Satt129; Satt147; Satt216; P10621B-2; Satt558;Satt266; Satt282; Satt537; Satt506; P13072A-1; Satt582; Satt389;Satt461; Satt514; Satt464; Satt543; P13074A-1; P10624A-1; Satt573;Satt598; Satt204; Satt263; Satt491; Satt602; Satt151; Satt355; Satt452;Satt146; Satt193; Satt569; Satt343; Satt586; Satt423; Satt348; Satt595;P10782A-1; P3436A-1; Satt334; Satt510; Satt144; Satt522; P9026A-1;P10646A-1; P5219A-1; P7659A-2; Satt570; Satt356; Satt130; Satt115;Satt594; Satt533; Satt303; Satt352; Satt566; Satt199; Satt503; Satt517;Satt191; SAT_(—)117; Satt353; Satt442; Satt279; Satt314; Satt181;Satt367; Satt127; Satt270; Satt440; P10640A-1; Satt249; SCT 065;Satt596; Satt280; Satt406; Satt380; Satt183; Satt529; Satt431; Satt742;Satt102; Satt240; P10618A-1; Satt523; Satt398; Satt497; Satt166;Satt448; Satt373; Satt513; P12394A-1; Satt590; Satt220; Satt536;Satt175; P10615A-1; Satt250; Satt346; Satt336; P13069A-1; P5467A-1;Satt584; SAT_(—)0844; Satt387; Satt339; P12396A-1; Satt487; Satt759;Satt347; Satt420; Satt576; Satt550; Satt262; Satt473; Satt477; Satt581;P11070A-1; Satt153; Satt243; P8230A-1; P10623A-1; P10632A-1; P12391A-1;P12392A-1; P13560A-1; P13561A-1; p2481A-1; SAC1677; Satt040; Satt109;Satt111; Satt176; Satt219; Satt299; and Satt512. The genetic elementcontributing to yield can be localized to any portion of the chromosomesegment defined by these molecular markers. For example, a gene encodinga function leading to improved yield can reside within the chromosomesegment at a distance of less than 1 CM (at a distance resulting inrecombination in fewer than 1% of mitotic events), or at any distancebetween about 1 CM and 10 CM, e.g., values of approximately 2 CM, 5 CM,4 CM, 5 CM, 6 CM, 7 CM, 8 CM, or 9 CM, or any value between about 0 CMand about 10 CM. Thus, a chromosome segment is defined as a contiguouslength of chromosome extending up to 10 CM in either direction from adesignated marker, and including the portion of the chromosome thatundergoes crossover with the marker locus at a frequency of no greaterthan 10%. In many cases, the chromosome segment of interest, is in facta continuous length extending less than 10 CM, i.e., a length ofchromosome that undergoes crossover with the marker at a frequency ofless than 10%, e.g., 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less, or anyvalue therebetween. Alternatively, physical distances may be utilized todetermine the aforementioned chromosome segments using a conversionfactor of 1 (one) Mbp (one million base pairs) for 1 (one) CM of mapdistance or 1% (one percent) crossover frequency. The actual conversionfactor in soybean ranges from about 100 Kbp to close to 1 Mbp dependingon the chromosome region. Accordingly, the arbitrary but reasonableconversion factor of 1 Mbp is to be understood in the context of thepresent invention.

The exemplary favorable alleles of marker loci enumerated herein havebeen shown, e.g., by breeding bias analysis, to be associated withgenetic elements contributing to improved yield in various lines ofsoybeans. One of skill will appreciate, that this is an empiricalassociation, and that a particular marker locus (and the designatedfavorable alleles thereof) are used to identify a correspondingchromosome segment having a genetic element that contributes to yield.However, because a marker locus is not necessarily, or even typically,identical to the genetic element which results in enhanced yield, insome percentage of crosses, genetic recombination will occur between theenumerated favorable allele and the genetic element contributing toyield. This does not negate the importance of either the genetic elementor the identification of the marker locus and exemplary favorablealleles. Any such recombination events can be detected and subsequentmarker assisted selection can be performed using the allele determinedto be in coupling linkage phase with the favorable genetic element asdiscussed in more detail in EXAMPLE 4: SEGREGATION FOR YIELD IN NEARISO-GENIC SUBLINES, in the context of detecting residual polymorphismsassociated with yield in near iso-genic sublines, and in EXAMPLE 5:ALLELE CONFIRMATION USING RANDOM CROSSES.

In the context of the present invention, the allelic form of multiplechromosome segments is determined, whether the purpose is to define thegenotype of a soybean or for marker assisted selection (MAS). In mostcircumstances, it is desirable to ascertain the allelic form for a largeproportion, e.g., all, of the chromosome segments known to contribute toyield in a particular geographic region. However, in some circumstances,particularly when the parents are known to share particular favorablealleles, a subset of the chromosome segments can be employed, reducingtime and cost, without losing efficiency. Thus, in the context of thepresent invention it is common to assess at least 10%, typically betweenat least 10% and about 90% of the relevant chromosome segments, e.g., bydetermining the allelic form of the marker loci specified in Tables 3through 12. Commonly, at least about 25%, frequently at least about 50%,often at least about 75% or more of the allelic forms are determined.For example, at least about 80%, 85%, 90%, 95%, 97%, 98%, 99%.Accordingly, it is generally desirable to determine the allele at amajority of the loci shown to be relevant to agronomic performance in aparticular geographic region or growing environment. In one favorableembodiment, the allelic form of a set of markers defining a set ofchromosome segments consisting essentially of the markers shown to berelevant to agronomic performance in a particular region are determined.A set of markers is deemed to consist essentially of the markersrelevant in a particular region if it includes a sufficient number ofmarkers to prevent a diminishing yield phenotype or to prevent adecrease in efficiency of selection when the set of markers is used formarker assisted selection. Typically such a set will include at leastabout 90%, 95%, 97%, 98%, 99% or more of the markers. That is, no morethan 10% of the markers shown to be relevant in a particular region willbe omitted, e.g., no more than 5%, 3%, 2%, or 1% of the allelic forms ofthe relevant markers will be left undetermined.

Typically, the marker loci evaluated are shown to correlate withsuperior agronomic performance with a significance of greater that 95%.Often the marker loci are selected that exhibit a significance ofgreater than 99% within a geographic region of interest. If desired, theallelic form of multiple marker loci within the same chromosome segmentcan be determined. Frequently, the allelic form of a single markerrepresenting the chromosome segment is determined.

Identification of Residual Polymorphism

The marker loci identified by Breeding Bias are associated with geneticelements contributing to increased yield in soybean. Although particularalleles of the marker loci have been found to be statisticallycorrelated with increased yield according to the Breeding Bias analysis,alleles at one or more marker loci are typically not fixed within anelite line or among progeny derived from a progenitor soybean selectedfrom an elite strain of germplasm. Indeed, such residual polymorphismsprovide valuable genetic variation from which improved sublines can beselected. By identifying particular marker loci from among the set ofloci shown to be associated with genetic elements contributing to yield,the efficiency of selection can be improved.

To this end, marker loci with one or more segregating alleles in apopulation of progeny derived from an elite progenitor are evaluated toidentify the specific allele correlating with increased yield in thepopulation of progeny. Detailed descriptions of exemplary methods forconfirming the favorable allele in a population of progeny are providedin EXAMPLES 4 and 5.

Definition and Identification of Target Germplasm

The marker loci of the present invention can be used as proxies for locicontributing to improved yield to define the theoretically optimal or“target” soybean genotype. Accordingly, alleles of marker lociidentified by Breeding Bias, and confirmed to correlate with increasedyield in a given environmental or geographic context can be used todefine or identify a soybean plant with superior agronomic performance.While existing elite lines include favorable alleles at numerous markerloci (and corresponding functional loci contributing to yield), none ofthe existing elite lines yet incorporates all of the genetic elementscontributing to the ideal soybean genotype. Indeed, prior to the markersof the present invention, it would not have been possible to predict theideal soybean genotype (or define a target genotype) with respect toyield. Thus, a feature of the invention is the definition of a targetsoybean genotype for superior agronomic performance. Any soybean planthaving an increased number of favorable allelic forms of chromosomesegments defined according to the markers of the invention, i.e., thatmore closely approximates the target soybean genotype than existingelite lines, constitutes a feature of the present invention.

The markers and methods described herein provide the means foridentifying and developing soybean plants having the target soybeangenotype for superior agronomic performance and genomes having increasednumbers of favorable allelic forms of the relevant chromosome segmentsrelative to existing elite lines, and or relative to either parentgiving rise to the soybean plant genome. Accordingly, the methods of theinvention can be used to produce compositions, including whole plants,plant organs, seeds, and isolated genetic constituents, e.g., includingthe entire chromosomal complement of the genome, or a subset thereof,such as an individual chromosome or chromosome fragment (all of whichare collectively described by the term “soybean plant genome”) with anincreased number of genetic elements contributing to yield. Methods forseparating and isolating genomes or individual chromosomes are wellknown in the art and include, e.g., flow cytometry and pulse field gelelectrophoresis. Replicates of the soybean plant genome are alsoencompassed within the meaning of the term soybean plant genome.Replicates are identical or substantially identical (i.e., a mutantarising from mitotic division from the same progenitor cell) to theinitial soybean plant genome. Replicates can be created, for example, byyeast or bacterial artificial chromosomes or any of a variety of nucleicacid vectors or replication methods such as PCR (polymerase chainreaction).

A soybean plant genome(s) of the present invention can be from anindividual soybean plant. As used herein, the term “plant” includeswhole plants, plant organs (e.g., leaves, stems, roots, etc.), seeds,suspension cultures, embryos, meristematic regions, callus tissue,leaves, roots, shoots, gametophytes, sporophytes, pollen, andmicrospores.

Marker Assisted Selection and Breeding

The ultimate goal of any breeding program is to combine as manyfavorable alleles as possible into elite varieties of germplasm that aregenetically superior (with respect to one or more agronomic traits) totheir ancestors. The markers provided herein identify chromosomesegments, i.e., genomic regions, and alleles (allelic forms) that havebeen favored by long-term selection for yield. Accordingly, thesemarkers can be used for marker assisted selection of soybean plants withsuperior agronomic performance. For example, in a cross between parentsthat complement favorable alleles at the target loci, progeny can beselected that include more favorable alleles than either parent. Suchprogeny are predicted to be phenotypically superior to either parent asillustrated in Table 2.

Marker assisted selection (MAS), employing the markers of the presentinvention, and the chromosome segments they identify are useful in thecontext of a soybean breeding program to increase efficiency in yieldimprovements. Phenotypic screening for a trait of interest, such asyield, for large numbers of samples can be expensive, as well as timeconsuming. In addition, phenotypic screening alone is often unreliabledue to the effects of epistasis and non-genetic (e.g., environmental)contributions to the phenotype. MAS offers the advantage over fieldevaluation that it can be performed at any time of year regardless ofthe growing season or developmental stage. In addition, MAS facilitatesevaluation of organisms grown in disparate regions or under differentconditions.

A breeder of ordinary skill, desiring to breed soybean plants withincreased yield, can apply the methods for MAS described herein, using,e.g., the exemplary markers provided herein or linked markers localizedto the chromosome segments identified by markers provided in Tables 3through 12, to derive soybean lines with, superior agronomicperformance.

Genetic marker alleles, e.g., the exemplary markers provided Tables 3through 12, linked markers, QTL, identifying the chromosome segmentsencompassing genetic elements that are important for yield, are used toidentify plants that contain a desired genotype at one or more loci, andthat are expected to transfer the desired genotype, along with a desiredphenotype to their progeny. Marker alleles (or QTL alleles) can be usedto identify plants that contain a desired genotype at one locus, or atseveral unlinked or linked loci (e.g., a haplotype), and that would beexpected to transfer the desired genotype, along with a desiredphenotype to their progeny. Similarly, by identifying plants lacking thedesired allele, plants with an undesirable phenotype, e.g., plants withpoor yield, can be identified, and, e.g., eliminated from subsequentcrosses. It will be appreciated that for the purposes of MAS, the termmarker can encompass both marker and QTL loci as both can be used toidentify plants with a desired phenotype.

For example, MAS can be used to develop lines or strains of soybean andsoybean germplasm with superior agronomic performance by identifyingfavorable allelic forms of chromosome segments shown to be important,e.g., that include a genetic element, for yield. Favorable alleles ofmarkers defining the chromosome segments of interest, e.g., Satt684,Satt165, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155,Satt385, Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480,Satt632-TB, Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409,Satt228, Satt429, Satt426, Satt509, SAT_(—)261, Satt197, Satt519,Satt597, SCT_(—)026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1,Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556, Satt272,Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361, P10639A-1,Satt661-TB, Satt190, SAT_(—)311-DB, Satt338, Satt227, Satt640-TB,Satt422, Satt457, Satt457, Satt557, Satt319, SAT_(—)142-DB, Satt460,P13073A-1, Satt307, SCT_(—)028, Satt433, Satt357, Satt321, Satt267,Satt383, Satt295, Satt203, Satt507, SAT_(—)110, P10620A-1, Satt129,Satt147, Satt216, SAT_(—)351, P1062113-2, Satt701, Satt634, Satt558,Satt266, Satt282, Satt537, Satt506, Satt546, P13072A-1, Satt582,Satt389, Satt461, Satt311, Satt514, Satt464, Satt662, Satt543, Satt186,Satt413, Satt672, P13074A-1, P10624A-1, Satt573, Satt598, Satt204,Satt263, Satt491, Satt602, Satt151, Satt355, Satt452, SAT_(—)273-DB,Satt146, Satt193, Satt569, Satt343, Satt586, Satt040, Satt423, Satt348,Satt595, P10782A-1, P3436A-1, P10598A-1, Satt334, Satt510, Satt510,Satt144, Satt522, Satt522, P9026A-1, P10646A-1, P5219A-1, P7659A-2,Satt570, Satt356, Satt130, Satt115, Satt594, Satt533, Satt303, Satt352,Satt566, Satt199, Satt503, Satt517, Satt191, SAT_(—)117, Satt353,Satt442, Satt279, Satt314, Satt142, Satt181, Satt367, Satt127, SCTT012,Satt270, Satt292, Satt440, P10640A-1, Satt249, SAG1223, SAC1699,SCT_(—)065, Satt596, Satt280, Satt406, Satt380, Satt183, Satt529,Satt431, Satt242, Satt102, Satt441, Satt544, Satt617, Satt240,P10618A-1, Satt475, Satt196, SAT_(—)301, Satt523, Satt418, Satt418,Satt398, Satt497, Satt284, Satt166, Satt448, Satt373, Satt513,P12394A-1, Satt590, Satt567, Satt220, SAG1048, Satt536, Satt175,Satt677, Satt680, P10615A-1, Satt551, Satt250, Satt346, Satt336,SAT_(—)330-DB, P13069A-1, P5467A-1, P5467A-2, Satt584, SAT_(—)084,P3050A-2, SAT_(—)275-DB, Satt387, Satt549, Satt660, Satt339, Satt2S5,Satt257, Satt358, P12396A-1, Satt487, Satt259, Satt259, Satt347,Satt420, Satt576, Satt550, Satt633, Satt262, Satt473, Satt477, Satt581,P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1, P10793A-1,P12391A-1, P13560A-1, P13561A-1, P13561A-1, P2481A-1, S60021-TB,S60048-TB, S60076-TB, S60148-TB, S60149-TB, 560201-TB, S60243-TB,S60326-T13, S60338-TB, S60350-TB, S60361-TB, S60422-TB, 560440-TB,S60446-TB, S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB,S60585-TB, S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055,Satt040, Satt108, Satt109, Satt111, Satt176, Satt176, Satt219, Satt299,and Satt512 as enumerated Tables 3 through 12 are detected in a genomicsample of a soybean plant.

For example, in a breeding program designed to produce soybeans withincreased yield in the Central United States growing region (e.g.,exemplified by the growing conditions in Iowa), markers can be selectedfrom the following set of markers: Satt642, Satt042, Satt364, Satt454,Satt526, Satt300, Satt591, Satt155, Satt385, Satt511, P12390B-1,Satt632-TB, Satt429, SAT_(—)261, Satt197, P10641A-1, Satt556, Satt534,P10638B-2, Satt399. Satt361, P10639A-1, Satt661-TB, Satt190,SAT_(—)311-DB, Satt338, Satt640-TB, Satt557, Satt319, SAT_(—)142-DB,Satt460, Satt433, Satt357, Satt321, Satt295, Satt203, Satt507, Satt129,Satt147, SAT_(—)51, P10621B-2, Satt558, Satt701, Satt634, Satt582,Satt389, Satt464, Satt662, Satt672, Satt573, Satt598, Satt263, Satt602,Satt151, SAT_(—)273-DB, Satt146, Satt193, Satt569, Satt176, Satt343,Satt586, Satt040, Satt595, P10782A-1, Satt334, Satt144, Satt522,Satt570, Satt356, Satt533, Satt199, Satt517, Satt191, SAT_(—)117,Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223, SAC1699,SAT_(—)065, Satt596, Satt406, Satt380, Satt183, Satt529, Satt242,Satt617, Satt240, SAT_(—)301, Satt418, Satt398, Satt497, Satt166,Satt448, Satt373, Satt513, P12394A-1, SAG1048, Satt536, Satt175,Satt677, Satt680, P10615A-1, Satt551, Satt346, Satt336, SAT_(—)330-DB,P13069A-1, P5467A-1, P5467A-2, SAT_(—)084, SAT_(—)275-D13, Satt660,Satt339, P12396A-1, Sat_(—)358, Satt487, Satt259, Satt420, Satt576,Satt633, Satt477, Satt581, Satt153, Satt243, P10793A-1, P12391A-1,P12392A-1, P13560A-1, P13561A-1, S60021-TB, S60048-TB, S60076-TB,S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB,S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB, S60505-TB,S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB,S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt040, Satt111,Satt176, Satt219 and Satt299, such as: Satt684, Satt042, Satt364,Satt454, Satt526, Satt300, Satt591, Satt155, Satt385, Satt632-TB,Satt429, SAT-_(—)261, P10641A-1, Satt556, 1P1063813-2, Satt399, Satt361,Satt66′-TB, Satt190, SAT_(—)311-DB, Satt338, Satt640-TB, Satt557,Satt319, SAT_(—)142-DB, Satt321, Satt203, Satt129, Satt147, SAT_(—)351,P10621B-2, Satt701, Satt634, Satt582, Satt389, Satt464, Satt662,Satt672, Satt573, Satt598, Satt263, Satt151, SAT_(—)273-DB, Satt146,Satt193, Satt569, Satt343, Satt586, Satt040, Satt595, Satt334, Satt144,Satt522, Satt570, Satt356, Satt199, Satt517, Satt191, Sat_(—)117,Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223, SAC1699,Sat_(—)065, Satt596, Satt406, Satt380, Satt183, Satt529, Satt242,Satt617, Satt240, SAT_(—)301, Satt418, Satt398, Satt497, Satt166,Satt448, Satt373, Satt513, P12394A-1, SAG1048, Satt536, Satt175,Satt677, Satt680, P10615A-1, Satt551, SAT_(—)330-D13, P13069A-1,P5467A-1, P5467A-2, SAT_(—)084, SAT_(—)275-DB, Satt660, Satt339,Satt358, Satt487, Satt487, Satt420, Satt576, Satt633, Satt581, Satt153,Satt243, P10793A-1, P13560A-1, P13561A-1, S60021-TB, S60048-TB,S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,S60338-TB, 560350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt111,Satt219 and Satt299. The following exemplary markers represent the bestmarkers for selection for yield in the Central Region in each chromosomeregion: Satt684, Satt526, Satt591, Satt385, Satt632-TB, Satt429,SAT-_(—)261, P10641A-1, Satt556, P10638B-2, Satt190, SAT_(—)311-DB,Satt338, Satt640-TB, Satt557, SAT_(—)142-DB, Satt321, Satt203, Satt129,SAT_(—)351, Satt701, Satt582, Satt389, Satt464, Satt672, Satt598,Satt343, Satt595, Satt334, Satt144, Satt522, Satt570, Satt356, Satt199,Sat_(—)117, Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223,Sat_(—)065, Satt529, Satt242, Satt617, SAT_(—)301, Satt398, Satt497,Satt166, Satt373, SAG1048, Satt680, P10615A-1, SAT_(—)330-DB, P13069A-1,SAT_(—)275-DB, Satt339, Satt487, Satt420, Satt581 and Satt153.Comparable lists of markers can be compiled from Tables 3 through 12 atthe discretion of the practitioner based on the desired growing region.

A soybean plant so identified can be utilized in a plant breedingprogram to develop lines with improved yield. Similarly, the detectionof favorable (or conversely, non-favorable) allelic forms of thechromosome segments can be used to trace the flow of alleles in asoybean plant pedigree to ensure that the desired complement of allelesare included or excluded in the resulting soybean plant(s).

After a desired phenotype and a polymorphic chromosomal locus, e.g., amarker locus or QTL, are determined to segregate together (i.e., aredetermined to be in linkage disequilibrium), alleles corresponding tothe desired phenotype are selected. In brief, a nucleic acidcorresponding to the marker nucleic acid is detected in a biologicalsample from a plant to be selected. This detection can take the from ofhybridization of a probe nucleic acid to a marker, e.g., usingallele-specific hybridization, Southern analysis, northern analysis, insitu hybridization, hybridization of primers followed by PCRamplification of a product including the marker, or the like. A varietyof procedures for detecting markers are described herein, e.g., in thesection entitled “DETECTION OF MARKER LOCI.” After the presence (orabsence) of a particular marker in the biological sample is verified,the plant is selected and, optionally, crossed to produce progenyplants.

When a population is segregating for multiple loci affecting one ormultiple traits, e.g., multiple loci involved in resistance to singledisease, or multiple loci each involved in resistance to differentdiseases, the efficiency of MAS compared to phenotypic screening becomeseven greater because all the loci can be processed in the lab togetherfrom a single sample of DNA. Thus, use of marker information for each ofthe traits in the breeding process is facilitated.

It will be appreciated that plants positive for a marker of theinvention can be selected and crossed according to any breeding protocolrelevant to the particular breeding program. Accordingly, progeny can begenerated from a selected plant by crossing the selected plant to one ormore additional plants selected on the basis of the same marker or adifferent marker, e.g., a different marker correlating with superioragronomic performance, or a different phentoype of interest, e.g.,resistance to a particular disease. Alternatively, a selected plant canbe back crossed to one or both parents. Backcrossing is usually done forthe purpose of introgressing one or a few loci from a donor parent,e.g., a donor parent comprising exotic germplasm, into an otherwisedesirable genetic background from the recurrent (typically, an elite)parent. The more cycles of backcrossing that are performed, the greaterthe genetic contribution of the recurrent parent to the resultingvariety. A selected plant can also be outcrossed, e.g., to a plant orline not present in its genealogy. Such a plant can be selected fromamong a population subject to a prior round of analysis, or may beintroduced into the breeding program de novo. A plant positive for adesired marker can also be self-crossed (“selfed”) to create a truebreeding line with the same genotype.

In some instances, even if a marker is close enough to a QTL to detectbreeding bias, the marker may not be close enough for reliable MAS. Ifsuch a marker is far enough away from the QTL of interest, there may becrossing over between the marker and the QTL leading to repulsion phaselinkages (in the elite population) between the marker allele that wasoriginally linked in coupling to the favorable QTL allele. With time,the marker locus and the linked QTL locus could reach “linkageequilibrium” and this will prevent the use of that marker for reliableselection of the favorable QTL allele. For highly heritable traits,linkage phase between marker and QTL in any given parent can be easilydetermined through MAS followed by phenotypic characterization. However,linkage phase determination for traits of low heritability (e.g., yield)is much more difficult. In fad, the effects of single loci on yield maybe extremely difficult to measure even with highly replicated fieldtests. In addition, if yield genes were highly heritable, the continuousselection for this trait would have “fixed” all of the favorable allelesquickly and yield progress would have previously reached a plateau.Since steady yield progress continues in soybean, it does not appearthat a “yield plateau” has been reached yet (Specht et al., 1999, CropScience 39:1560-1570). Therefore, many of the favorable alleles at themajor yield loci in soybean are not yet fixed within the elitepopulation. The question remains whether the existing marker loci arestill in original linkage phase with the QTL loci. Fortunately, theresults of breeding bias disclosed here can be used to solve the problemof “imperfect” yield gene markers several ways: a) linkage phase and QTLeffect determination within specific crosses prior to MAS, b) use offlanking markers to predict which markers are still linked in coupling,and c) alternating cycles of MAS and phenotypic selection.

Linkage Phase and QTL Effect Determination within Specific Crosses:

Because elite soybean lines are highly related, a relatively small setof elite lines contains most of the favorable alleles that exist withinthe entire elite population. Therefore, determining the linkage phasebetween marker and QTL alleles at the loci identified as important foryield can be accomplished to further increase efficiency of MAS in thecontext of a soybean breeding program. Progeny from parents that areboth high yielding and polymorphic for many of the target marker lociare assessed for yield in a small number of field locations. Usingorthoganol comparisons, one locus at a time, predictions concerningcorrelations between marker alleles and phenotype can be made. Forexample if 40 random homozygous progeny from a cross that is segregatingat 10 of the target loci are field tested, on average, 20 of the progenywill be homozygous for one of the parental alleles and 20 will behomozygous for the other parental allele. One can then pool thereplicated yield data for lines containing the same marker allele anddetermine if the yield of said group is statistically different from theyield of lines containing the alternate marker allele. If so, then thismarker should be effective for selection within that cross. Thiscomparison is then done separately for each of the 10 segregating markerloci to determine which set of those 10 markers should be effective forselection within that cross.

Optionally, flanking markers can be used to predict which markers arelinked in coupling. By comparing the genotype of flanking markers thatare linked to the target marker in both elite lines and ancestors, onecan predict which haplotypes have been most conserved during selectionover many cycles. In the event that recombination has occurred betweenthe target marker and the genetic element contributing to yield, suchthat the desired genetic element and the linked target marker locus areno longer in coupling linkage phase, flanking markers can be utilized toidentify progeny with superior agronomic performance.

In addition, MAS and phenotypic selection can be alternated to insurethat the allele in coupling phase is detected. Markers identified bybreeding bias are employed for MAS as described above (with or withoutthe advantages of linkage phase determination within specific crosses)and replicated yield testing is performed on selected progeny. If enoughreplicates and environments are sampled, a reasonable measure of yieldphenotype can be obtained. Progeny that are confirmed as high yieldingcan be used as parents in the next cycle of MAS selection. By screeningrelated populations according to this method, the population will movetoward fixation of the favorable QTL alleles even if the favorablemarker allele is not always in coupling with the QTL allele.Alternatively, residual polymorphisms at the marker loci describedherein can be detected in near iso-genic lines, and the marker allele incorresponding to increased yield can be validated.

Introgression of Favorable Alleles

More Efficient Backcrossing of Specific Genes into Elite Lines

One application of MAS, in the context of the present invention is touse the “yield gene” markers to increase the efficiency of aintrogression or backcrossing effort. In typical marker assistedbackcrossing of a specific gene(s) from a donor source to an elitegenetic background, one selects among backcross progeny for the donortrait and then uses markers to reconstitute as much of the elitebackground's genome as possible. Prior to the present invention, themarkers used to identify the elite background were of unknown function,and many of the markers commonly used may be selecting for parts of theelite genome that do not actually contribute to high yield. Similarly,prior to the present invention, the major loci that contribute to yieldwere largely unknown, so the entire elite genome of the recurrent parentwas selected for with the hopes of including all of the favorablealleles that it contained. However, the markers identified by breedingbias can be used to identify only those parts of the elite genome thatare most significant with respect to yield. These markers can be used toconcentrate backcrossing efforts on the most important parts of theelite genome. The fewer markers needed, the higher the probability ofrecapturing the elite phenotype quickly.

Thus, the markers and methods of the present invention can be utilizedto guide marker assisted selection or breeding of soybean varieties withthe desired complement (i.e., set) of allelic forms of chromosomesegments associated with superior agronomic performance. Each of thedisclosed alleles can be introduced into a soybean line viaintrogression, i.e., by means of traditional breeding (or introduced viatransformation, or both) to yield a soybean plant with superioragronomic performance. The number of alleles associated with superioragronomic performance that can be introduced or be present in a soybeanplant of the present invention ranges from 1 to the number of allelesdisclosed herein, each integer of which is incorporated herein as ifexplicitly recited.

Exemplary soybean, lines including at least one (and typically severalor many) of the favorable allelic forms of the relevant chromosomesegments are provided in Table 1. Without intent to limit the invention,these include the elite soybean lines: 90A07, 90B11, 90B31, 90B43,90B72, 90B73, 91B01, 91B12, 91B33, 91B52, 91B53, 91B64, 91B91, 91B92,92B05, 92B12, 92B23, 92B38, 92B63, 92B74, 92B75, 92B84, 92B95, 93B01,93B11, 93B15, 93B25, 93B26, 93B41, 93B45, 93B46, 93B66, 93B67, 93B72,93B82, 93B84, 93B85, 93B86, 93B87, 94B01, 94B23, 94B24, 94B53, 94B54,94B73, 95B32, 95B33, 95B34, 95B53, 95B95, 95B96, 95B97, 96B21, 96B51,97B52, 97B61, A1395, A2835, A2943, A3127, A3242, A3431, A4009, A4138,A4415, A4595, A4715, A5403, A5560, A5843, A5885, A5979, A5980, A6297,BEDFORD, CM428, CX105, CX232, CX253, CX289, CX394C, CX469C, D00566D362,ESSEX, EX04C00, EX06A00, EX 10E01, EX13P01, EX13Q01, EX 15N01, EX16N00,EX16P01, EX22Y01, EX22Z01, EX39E00, FORREST, G3362, HS93-4118,HUTCHESON, KORADA, MO15733, MO400644-02, MO413735-11-52, MO501577-27-23,MO505469-61-89, MP39009, P1677, P9007, P9008, P9041, P9042, P9061,P9062, P9063, P9071, P9092, P9132, P9141, P9151, P9163, P91B2, P9203,P9233, P9244, P9273, P9281, P9305, P9306, P9321, P9341, P9392, P9395,P9481, P9482, P9492, P9521, P9552, P9561, P9584, P9591, P9592, P9594,P9631, P9641, PHARAOH, RA451, R01154R002, S0066, S03W4, S0880, S1550,S1990, S19T9, S20F8, S22C3, S24L2, S25J5, S32Z3, S33N1, S38T8, S3911,S4260, S42H1, S4385, S5960, S6189, S6262, ST0653, ST1073, ST1090,ST1970, ST2250, ST2488, ST2660, ST2688, ST2870, ST3171, ST3380, ST3630,ST3870, ST3883, TRACY, TRAILL, X9916, YB03E00, XB03F01, XB07E01,XB10D01, XB15M01, XB20M01, XB22R01, XB25W01, XB31C01, XB3313, XB34F01,XB35D, XB35W00, XB38A01, XB41M01, XB42J00, XB42M01, XB48H01, XB54K01,XB55J01, XB58P99, XB63D00, XB67A00, YB03G01, YB08D01, YB09F01, YB09G01,YB10E01, YB11D01, YB14H01, YB15K99, YB21F01, YB21G01, YB22S00, YB22V01,YB22W01, YB22X01, YB24Z01, YB25R99, YB25X00, YB25Y01, YB25Z01, Y1327X01,YB27Y01, YB28N01, YB29H01, YB29J01, YB30J01, YB30N01, YB30P01, YB31E01,YB32K01, YB33K01, YB34H01, YB35C01, YB36V00, YB39M01, YB40M01, YB40N01,YB41Q01, YB48L01, YB52J00, YB53E00, YB54H00, YB54J00, YB54L00, YB55H00,YB56E00, YB60N01, and YOUNG. These lines and progeny derived therefrom,as well as numerous additional elite lines, are conveniently utilized asbreeding material to develop novel lines with increased numbers offavorable allelic forms of chromosome segments involved in yield.

The present invention also extends to a method of making a progenysoybean plant and these progeny soybean plants, per se. The methodcomprises crossing a first parent soybean plant with a second soybeanplant and growing the female soybean plant under plant growth conditionsto yield soybean plant progeny. Methods of crossing and growing soybeanplants are well within the ability of those of ordinary skill in theart. Such soybean plant progeny can be assayed for the allelesassociated with superior agronomic performance and, thereby, the desiredprogeny selected. Such progeny plants or seed can be sold commerciallyfor soybean production, used for food, processed to obtain a desiredconstituent of the soybean, or further utilized in subsequent rounds ofbreeding. At least one of the first or second soybean plants is asoybean plant of the present invention in that it comprises at least oneof the allelic forms of the present invention such that the progeny arecapable of inheriting the allele. Conveniently, the first or secondsoybean plant line can be one of the elite lines of Table 1, or aderivative of such a line (i.e., a descendant or progenitor in thatline's pedigree), or any relative of these elite lines (such as anyelite line that was derived from the ancestors of these elite lines)that retains the same allelic form as that associated with superioragronomic performance. However, it will readily be recognized by one ofskill in the art that following characterization essentially any eliteline of soybean can be utilized.

Often, a method of the present invention is applied to at least onerelated soybean plant such as from progenitor or descendant lines in thesubject soybean plants pedigree such that inheritance of the desiredallele can be traced. The number of generations separating the soybeanplants being subject to the method of the present invention willgenerally be from 1 to 20, commonly 1 to 5, and typically 1, 2, or 3generations of separation, and quite often a direct descendant or parentof the soybean plant will be subject to the method (i.e., 1 generationof separation).

Incorporation of “Exotic” Germplasm while Maintaining HistoricalProgress

Genetic diversity is important for long term genetic gain in anybreeding program. With limited diversity, genetic gain will eventuallyplateau when all of the favorable alleles have been fixed within theelite population. The challenge is to incorporate diversity into theelite pool without losing the genetic gain that has already been madeand with the minimum possible investment. Breeding bias results providean indication of which genomic regions and which favorable alleles fromthe original ancestors have been selected for and conserved over time,facilitating efforts to incorporate favorable variation from exoticgermplasm sources (parents that are unrelated to the elite gene pool) inthe hopes of finding favorable alleles that do not currently exist inthe elite gene pool.

For example, the markers of the present invention can be used for MAS incrosses involving elite×exotic soybean lines by subjecting thesegregating progeny to MAS to maintain the major yield alleles that havealready been “fixed” by decades of selection and leave the rest of thegenome open for contribution from the exotic sources. This would be amuch more efficient system than conventional selection or MAS selectionof elite alleles for which we have no prior information.

If the donor parent has polymorphic alleles at the elite target loci aswell, the breeder can also relax the backcrossing selection intensity toallow variation at these loci to slip through. This provides theopportunity to see if an exotic line has something even more favorablethan what was in the elite gene pool at the elite target loci.

The methods of the present invention also address another limitation ofconventional backcrossing: that is, as the recurrent parent isreconstituted with increased cycles of backcrossing, potentiallyfavorable alleles from the donor parent are excluded along with theunfavorable alleles from the donor parent. By selectively reconstitutingthe recurrent parent's genotype at the loci that have been shown bybreeding bias to be important, one allows favorable alleles to beintroduced from the donor parent at other loci. This allows for thedonor parent to contribute exotic favorable alleles at loci that are notpart of the elite “target genotype.” This increases the chances oftransgressive segregation while still retaining the most critical partsof the recurrent parent's genome. If the donor parent has polymorphicalleles at the elite target loci as well, the breeder can also relax thebackcrossing selection intensity to allow variation at these loci toslip through and be tested in the context of a genome that isrepresentative of the elite gene pool.

Detection of Marker Loci

Tables 3 through 12 provide a set of markers and favorable allelesassociated with superior agronomic performance in a variety ofgeographic regions with a range of different growing environments. Eachof the markers identifies a chromosome segment that includes one or moregenetic elements, i.e., genes, that influences yield in soybeans. One ofskill in the art will appreciate that the markers provided and discussedherein are merely exemplary and that numerous other linked markers canbe identified based on genetic linkage and/or physical proximity on achromosome to the markers provided herein. Thus, the compositions andmethods of the present invention described herein are not intended to belimited only to the markers provided in Tables 3 through 12, but alsoinclude additional markers linked thereto. Additionally, while favorablealleles of the exemplary marker loci are disclosed herein, it will bereadily appreciated by those of skill in the art, that favorable allelesof additional loci linked to the marker loci described herein can bedetermined without undue experimentation and employed in thecompositions and methods of the present invention. Accordingly, anymarker locus linked to the markers described herein, and localized to achromosome segment identified by the markers of the invention, can alsobe used to identify that chromosome segment, and to define the genotypeof a soybean plant, or to select for favorable allelic forms of achromosome segment correlated with superior agronomic performance.

Although the specific DNA sequences which encode proteins are generallywell-conserved across a species, regions of DNA which are non-coding, orwhich encode proteins or portions of proteins which lack criticalfunction, tend to accumulate mutations, and therefore, are variablebetween members of the same species. Such regions provide the basis fornumerous molecular genetic markers. Markers identify alterations in thegenome, which can be insertions, deletions, point mutations,recombination events, or the presence and sequence of transposableelements. Many molecular or genetic markers have been characterized inplant species of interest, including soybean, and are known to those ofskill in the art. For example, a collection of genetic markers forsoybean is publicly available from Linkage Genetics (151 West 2200South, Suite C, Salt Lake City, Utah 84119, 801-975-1188).

Molecular markers can be detected by numerous methods, well-establishedin the art (e.g., allele specific hybridization (ASH) or other methodsfor detecting single nucleotide polymorphisms (SNP), amplified fragmentlength polymorphisms (AFLP), amplified variable sequences, randomlyamplified polymorphic DNA (RAPD), restriction fragment lengthpolymorphisms (RFLP), self-sustained sequence replication, simplesequence repeat (SSR), single-strand conformation polymorphisms (SSCP),and isozyme markers). While the exemplary markers provided in Tables 3through 12 are either SSR or SNP (ASH) markers, any of theaforementioned marker types can be employed in the context of theinvention to identify chromosome segments encompassing genetic elementthat contribute to superior agronomic performance.

The majority of genetic markers rely on one or more property of nucleicacids for their detection. For example, some techniques for detectinggenetic markers utilize hybridization of a probe nucleic acid to nucleicacids corresponding to the genetic marker. Hybridization formatsincluding but not limited to, solution phase, solid phase, mixed phase,or in situ hybridization assays. Among the earliest markers detected,restriction fragment length polymorphisms (RFLP), are detected byhybridizing a probe which is typically a sub-fragment (or a syntheticoligonucleotide corresponding to a sub-fragment) of the nucleic acid tobe detected to restriction digested genomic DNA. The restriction enzymeis selected to provide restriction fragments of at least two alternative(or polymorphic) lengths in different individuals, and will often varyfrom line to line. Determining a (one or more) restriction enzyme thatproduces informative fragments for each cross is a simple procedure,well known in the art. After separation by length in an appropriatematrix (e_g, agarose) and transfer to a membrane (e.g., nitrocellulose,nylon), the labeled probe is hybridized under conditions which result inequilibrium binding of the probe to the target followed by removal ofexcess probe by washing.

Nucleic acid probes to the marker loci can be cloned and/or synthesized.Detectable labels suitable for use with nucleic acid probes include anycomposition detectable by spectroscopic, radioisotopic, photochemical,biochemical, immunochemical, electrical, optical or chemical means.Useful labels include biotin for staining with labeled streptavidinconjugate, magnetic beads, fluorescent dyes, radiolabels, enzymes, andcolorimetric labels. Other labels include ligands that bind toantibodies labeled with fluorophores, chemiluminescent agents, andenzymes. Labeling markers is readily achieved such as by the use oflabeled PCR primers to marker loci.

The hybridized probe is then detected using, most typically byautoradiography or other similar detection technique (e.g.,fluorography, liquid scintillation counter, etc.). Examples of specifichybridization protocols are widely available in the art, see, e.g.,Berger, Sambrook, Ausubel, cited in the section entitled “GENERALMOLECULAR BIOLOGY REFERENCES.”

More specifically with respect to certain of the exemplary markers ofthe present invention, Allele-specific hybridization (ASH) technology isbased on the stable annealing of a short, single-stranded,oligonucleotide probe to a completely complementary single-strand targetnucleic acid. Detection is via an isotopic or non-isotopic labelattached to the probe.

For each polymorphism, two of more different ASH probes are designed tohave identical DNA sequences except at the polymorphic nucleotides ofmarkers comprising a single nucleotide polymorphism (SNP). Each probewill have exact homology with one allele sequence so that the range ofprobes can distinguish all the known alternative allele sequences. Eachprobe is hybridized to the target DNA. With appropriate probe design andhybridization conditions, a single-base mismatch between the probe andtarget DNA will prevent hybridization. In this manner, only one of thealternative probes will hybridize to a target sample that is homozygousor homogenous for an allele. Samples that are heterozygous orheterogeneous for two alleles will hybridize to both, of two alternativeprobes.

ASH markers are used as dominant markers where the presence or absenceof only one allele is determined from hybridization or lack ofhybridization by only one probe. The alternative allele may be inferredfrom the lack of hybridization. ASH probe and target molecules areoptionally RNA or DNA; the target molecules are any length ofnucleotides beyond the sequence that is complementary to the probe; theprobe is designed to hybridize with either strand of a DNA target; theprobe ranges in size to conform to variously stringent hybridizationconditions, etc.

PCR allows the target sequence for ASH to be amplified from lowconcentrations of nucleic acid in relatively small volumes. Otherwise,the target sequence from genomic DNA is digested with a restrictionendonuclease and size separated by gel electrophoresis. Hybridizationstypically occur with the target sequence bound to the surface of amembrane or, as described in U.S. Pat. No. 5,468,613, the ASH probesequence may be bound to a membrane.

In one embodiment, ASH data are obtained by amplifying nucleic acidfragments (amplicons) from genomic DNA using PCR, transferring theamplicon target DNA to a membrane in a dot-blot format, hybridizing alabeled oligonucleotide probe to the amplicon target, and observing thehybridization dots by autoradiography.

Other of the exemplary molecular markers provided herein are Simplesequence repeats (SSR). SSR markers take advantage of high levels ofdi-, tri-, or tetra-nucleotide tandem repeats within a genome.Dinucleotide repeats have been reported to occur in the human genome asmany as 50,000 times with n varying from 10 to 60 or more (Jacob et al.(1991) Cell 67:213. Dinucleotide repeats have also been found in higherplants (Condit and Hubbell (1991) Genome 34:66).

Briefly, SSR data is generated by hybridizing primers to conservedregions of the plant genome which flank the SSR sequence. PCR is thenused to amplify the dinucleotide repeats between the primers. Theamplified sequences are then electrophoresed to determine the size andtherefore the number of di-, tri-, and tetra-nucleotide repeats.

Amplified variable sequences refer to amplified sequences of the plantgenome which exhibit high nucleic acid residue variability betweenmembers of the same species, e.g., microsatellite sequences. Allorganisms have variable genomic sequences and each organism (with theexception of a clone) has a different set of variable sequences. Onceidentified, the presence of specific variable sequences can be used topredict phenotypic traits. Preferably, DNA from the plant serves as atemplate for amplification with primers that flank a variable sequenceof DNA. The variable sequence is amplified and then sequenced.

Randomly amplified polymorphic DNA (RAPD) markers are genomic sequencesamplified by PCR using a single short primer of arbitrary sequence atlow stringency. During amplification at low stringency a number of PCRproducts, some of which differ in length (and sequence) betweenindividuals, are generated from random locations throughout the genome.Unlike amplified variable sequences, no prior sequence information isrequired to identify RAPD markers.

In vitro amplification techniques are well known in the art. Examples oftechniques sufficient to direct persons of skill through such in vitromethods, including the polymerase chain reaction (PCR), the ligase chainreaction (LCR), Qβ-replicase amplification and other RNA polymerasemediated techniques (e.g., NASBA), are found in Berger, Sambrook andAusubel as well as Mullis et al. (1987) U.S. Pat. No. 4,683,202; PCRProtocols, A Guide to Methods and Applications (Innis et al., eds.)Academic Press Inc., San Diego Academic Press Inc. San Diego, Calif.(1990) (Innis); Arnheim & Levinson (Oct. 1, 1990) C&EN 36-47; TheJournal Of NIH Research (1991) 3, 81-94; (Kwoh et al. (1989) Proc. Natl.Acad. Sci. USA 86, 1173; Guatelli at al. (1990) Proc. Natl. Acad. Sci.USA 87, 1874; Lomell et al. (1.989) J. Chin. Chem 35, 1826; Landegren etal., (1988) Science 241, 1077-1080; Van Brunt (1990) Biotechnology 8,291-294; Wu and Wallace, (1989) Gene 4, 560; Barringer at al. (1990)Gene 89, 117, and Sooknanan and Malek (1995) Biotechnology 13: 563-564.Improved methods of cloning in vitro amplified nucleic acids aredescribed in Wallace at al., U.S. Pat. No. 5,426,039. Improved methodsof amplifying large nucleic acids by PCR are summarized in Cheng et al.(1994) Nature 369: 684, and the references therein, in which PCRamplicons of up to 40 kb are generated. One of skill will appreciatethat essentially any RNA can be converted into a double stranded DNAsuitable for restriction digestion, PCR expansion and sequencing usingreverse transcriptase and a polymerase. See, Ausubel, Sambrook andBerger.

Oligonucleotides for use as primers, e.g., in amplification reactionsand for use as nucleic acid sequence probes are typically synthesizedchemically according to the solid phase phosphoramidite triester methoddescribed by Beaucage and Caruthers (1981) Tetrahedron Lett. 22:1859, orcan simply be ordered commercially.

Alternatively, self-sustained sequence replication can be used toidentify genetic markers. Self-sustained sequence replication refers toa method of nucleic acid amplification using target nucleic acidsequences which are replicated exponentially in vitro undersubstantially isothermal conditions by using three enzymatic activitiesinvolved in retroviral replication: (1) reverse transcriptase, (2) RnaseH, and (3) a DNA-dependent RNA polymerase (Guatelli et al. (1990) ProcNatl Acad Sci USA 87:1874). By mimicking the retroviral strategy of RNAreplication by means of cDNA intermediates, this reaction accumulatescDNA and RNA copies of the original target.

Amplified restriction fragment polymorphisms or amplified fragmentlength polymorphisms (AFLP) can also be used as genetic markers (Vos etal, (1995) Nucl Acids Res 23:4407. The phrase “amplified restrictionfragment polymorphism” refers to selected restriction fragments, whichare amplified before or after cleavage by a restriction endonuclease.The amplification step allows easier detection of specific restrictionfragments. AFLP allows the detection large numbers of polymorphicmarkers and has been used for genetic mapping of plants (Becker et al.(1995) Mol Gen Genet. 249:65; and Meksem et al. (1995) Mol Gen Genet.249:74.

Single nucleotide polymorphism (SNP) are markers that consist of ashared sequence differentiated on the basis of a single nucleotide.Typically, this distinction is detected by differential migrationpatterns of an amplicon comprising the SNP on e.g., an acrylamide gel.In such cases the marker may also be referred to as a single-strandconformation polymorphism or SSCP. However, alternative modes ofdetection, such as hybridization, e.g., ASH, or RFLP analysis are notexcluded.

Alternatively, isozyme markers are employed as genetic markers. Isozymesare multiple forms of enzymes that differ from one another in theiramino acid, and therefore their nucleic acid sequences. Some isozymesare multimeric enzymes containing slightly different subunits. Otherisozymes are either multimeric or monomeric but have been cleaved fromthe proenzyme at different sites in the amino acid sequence. Isozymescan be characterized and analyzed at the protein level, oralternatively, isozymes that differ at the nucleic acid level can bedetermined. In such cases any of the nucleic acid based methodsdescribed herein can be used to analyze isozyme markers.

In alternative embodiments, in silico methods can be used to detect themarker loci. For example, the sequence of a nucleic acid comprising themarker can be stored in a computer. The desired marker locus sequence orits homolog can be identified using an appropriate nucleic acid searchalgorithm as provided by, for example, in such readily availableprograms as BLAST.

Integrated Systems/Computer Assisted Methods

In some embodiments, the present invention includes an “integratedsystem” including an electronic means of storing or transmittingcomputer readable data representing or designating the allelic formsdetermined by the method of the present invention. The computer readablemedia includes cache, main, and storage memory and other electronic datastorage means for storage of computer code. Data representing theallelic forms determined by the method of the present invention can alsobe electronically transmitted in a computer data signal embodied in atransmission medium over a network such as an intranet or internet orcombinations thereof.

The phrase “integrated system” in the context of this invention refersto a system in which data entering a computer corresponds to physicalobjects or processes external to the computer, e.g., a marker allele,and a process that, within a computer, causes a physical transformationof the input signals to different output signals. In other words, theinput data, e.g., amplification of a particular marker allele istransformed to output data, e.g., the identification of the allelic formof a chromosome segment. The process within the computer is a set ofinstructions, or “program,” by which positive amplification orhybridization signals are recognized by the integrated system andattributed to individual samples as a genotype. Additional programscorrelate the identity of individual samples with phenotypic values ormarker alleles, e.g., statistical methods. In addition there arenumerous e.g., C/C++ programs for computing, Delphi and/or Java programsfor GUI interfaces, and productivity tools (e.g., Microsoft Excel and/orSigmaPlot) for charting. Other useful software tools in the context ofthe integrated systems of the invention include statistical packagessuch as SAS, Genstat, Matlab, Mathematica, and S-Plus and geneticmodeling packages such as QU-GENE. Furthermore additional programminglanguages such as Fortran and the like are also suitably employed in theintegrated systems of the invention.

For example, marker allele values assigned to a population of progenydescending from crosses between elite lines are recorded in a computerreadable medium, thereby establishing a database corresponding allelicforms with unique identifiers for each member of the population ofprogeny. Any file or folder, whether custom-made or commerciallyavailable (e.g., from Oracle or Sybase) suitable for recording data in acomputer readable medium is acceptable as a database in the context ofthe present invention. Data regarding genotype for one or more molecularmarkers, e.g., ASH, SSR, RFLP, RAPD, AFLP, SNP, isozyme markers or othermarkers as described herein, are similarly recorded in a computeraccessible database. Optionally, marker data is obtained using anintegrated system that automates one or more aspects of the assay (orassays) used to determine marker(s) genotype. In such a system, inputdata corresponding to genotypes for molecular markers are relayed from adevice, e.g., an array, a scanner, a CCD, or other detection devicedirectly to files in a computer readable medium accessible to thecentral processing unit. A set of instructions (embodied in one or moreprograms) encoding the statistical models of the invention is thenexecuted by the computational device to identify correlations betweenyield data and marker genotypes. Typically, the integrated system alsoincludes a user input device, such as a keyboard, a mouse, atouchscreen, or the like, for, e.g., selecting files, retrieving data,etc., and an output device (e.g., a monitor, a printer, etc.) forviewing or recovering the product of the statistical analysis.

Thus, in one aspect, the invention provides an integrated systemcomprising a computer or computer readable medium comprising set offiles and/or a database with at least one data set that corresponds togenotypes for genetic markers. The system also includes a user interfaceallowing a user to selectively view one or more databases. In addition,standard text manipulation software such as word processing software(e.g., Microsoft Word™ or Corel Wordperfect™ and database or spreadsheetsoftware (e.g., spreadsheet software such as Microsoft Excel™, CorelQuattro Pro™, or database programs such as Microsoft Access™ orParadox™) can be used in conjunction with a user interface (e.g., a GUIin a standard operating system such as a Windows, Macintosh, Unix orLinux system) to manipulate strings of characters.

The invention also provides integrated systems for sample manipulationincorporating robotic devices as previously described. A robotic liquidcontrol armature for transferring solutions (e.g., plant cell extracts)from a source to a destination, e.g., from a microtiter plate to anarray substrate, is optionally operably linked to the digital computer(or to an additional computer in the integrated system). An input devicefor entering data to the digital computer to control high throughputliquid transfer by the robotic liquid control armature and, optionally,to control transfer by the armature to the solid support is commonly afeature of the integrated system.

Integrated systems for molecular marker analysis of the presentinvention typically include a digital computer with one or more ofhigh-throughput liquid control software, image analysis software, datainterpretation software, a robotic liquid control armature fortransferring solutions from a source to a destination operably linked tothe digital computer, an input device (e.g., a computer keyboard) forentering data to the digital computer to control high throughput liquidtransfer by the robotic liquid control armature and, optionally, animage scanner for digitizing label signals from labeled probeshybridized, e.g., to expression products on a solid support operablylinked to the digital computer. The image scanner interfaces with theimage analysis software to provide a measurement of e.g.,differentiating nucleic acid probe label intensity upon hybridization toan arrayed sample nucleic acid population, where the probe labelintensity measurement is interpreted by the data interpretation softwareto show whether, and to what degree, the labeled probe hybridizes to alabel. The data so derived is then correlated with sample identity, todetermine the identity of a plant with a particular genotype(s) forgenetic markers, e.g., to facilitate marker assisted selection ofsoybean plants with favorable allelic forms of chromosome segmentsinvolved in agronomic performance.

Optical images, e.g., hybridization patterns viewed (and, optionally,recorded) by a camera or other recording device (e.g., a photodiode anddata storage device) are optionally further processed in any of theembodiments herein, e.g., by digitizing the image and/or storing andanalyzing the image on a computer. A variety of commercially availableperipheral equipment and software is available for digitizing, storingand analyzing a digitized video or digitized optical image, e.g., usingPC (Intel x86 or pentium chip-compatible DOS™, OS2™ WINDOWS™, WINDOWSNT™ or WINDOWS95™ based machines), MACINTOSH™, LINUX, or UNIX based(e.g., SUN™ work station) computers.

Identification of Additional Markers and QTL Associated With Yield

Nucleic acids isolated from the chromosome segments of the presentinvention, e.g., nucleic acids corresponding to additional marker loci,nucleic acids corresponding to genetic elements contributing to superioragronomic performance, are within the scope of the present invention.For example, in the rare instance in which the markers and alleles ofthe present invention are not well suited for MAS, the markers can,nonetheless, be used to identify additional linked markers that are,e.g., closer to, or within, the QTL of interest. Based on the markersdisclosed herein, improvement in the efficiency of MAS can be obtainedby saturating the relevant chromosome segments with numerous linkedmarkers. The breeding bias analysis can be repeated on all markerswithin the region to identify those having the highest statisticalsignificance. MAS can be practiced with all markers in the regionfollowed by phenotypic characterization (i.e., of yield) to determinewhich markers are most efficient.

Additional markers can be identified in a variety of ways. For example,additional markers can be identified by evaluating publicly or privatelyavailable markers that have been mapped to the genomic region(s) ofinterest, including candidate markers known to encode proteins that are,at least theoretically, likely to be related to yield. Alternatively,unmapped markers can be evaluated to determine which markers map to thesame genomic region via independent mapping studies.

A theoretically optimal marker can be obtained by isolating geneticelements contributing to superior agronomic performance, such as codingregions giving rise to expression products that influence yield.Identification of a QTL underlying superior agronomic performance can beaccomplished by anchoring the marker to a physical DNA map and thenprogressing upstream and downstream to identify coding sequences, i.e.,by “positional gene cloning.”

Positional gene cloning uses the physical proximity of a genetic marker(such as a marker provided in Tables 3 through 12) to identity a clonedchromosomal fragment that includes a nucleic acid of interest, e.g., aQTL contributing to superior agronomic performance. Clones of nucleicacids linked to the markers of the invention have a variety of uses,including as additional genetic markers to define the chromosomesegments of the invention and for use in marker assisted selection(MAS). Markers which are adjacent to an open reading frame (ORF) canhybridize to a DNA clone, thereby identifying a clone on which an ORFassociated with a trait contributing to yield is located. If the markeris more distant, a fragment containing the open reading frame isidentified by successive rounds of screening and isolation of cloneswhich together comprise a contiguous sequence of DNA, a “contig.”Protocols sufficient to guide one of skill through the isolation ofclones associated with linked markers are found in, e.g., in thereferences cited in the section entitled “GENERAL MOLECULAR BIOLOGYREFERENCES” below.

An isolated chromosome fragment can be produced by such well knownmethods as digesting chromosomal DNA with one or more restrictionenzymes, or by amplifying a chromosomal region in a polymerase chainreaction (PCR), or alternative amplification reaction. The digested oramplified fragment is typically ligated into a vector suitable forreplication, e.g., a plasmid, a cosmid, a phage, an artificialchromosome, or the like, and, optionally expression, of the insertedfragment.

Such chromosome segments can be utilized to identify homologous nucleicacids, e.g., in other lines or species, and/or can be used in theproduction of transgenic plants with desirable phenotypic attributesrelated to agronomic performance. A chromosome segment comprising anucleic acid contributing to increased yield is isolated, e.g., clonedvia positional cloning methods outlined above. A chromosome segment cancontain one or more ORFs associated with the desired phenotypic trait,and can be cloned on one or more individual vectors, e.g., depending onthe size of the chromosome interval.

It will be appreciated that numerous vectors are available in the artfor the isolation and replication of the nucleic acids of the invention.For example, plasmids, cosmids and phage vectors are well known in theart, and are sufficient for many applications (e.g., in applicationsinvolving insertion of nucleic acids ranging from less than 1 to about20 kilobases (kb). In certain applications, it is advantageous to makeor clone large nucleic acids to identify nucleic acids more distantlylinked to a given marker, or to isolate nucleic acids in excess of 10-20kb, e.g., up to several hundred kilobases or more, such as the entireinterval between two linked markers, i.e., up to and including one ormore centimorgans (CM), linked to markers as identified herein. In suchcases, a number of vectors capable of accommodating large nucleic acidsare available in the art, these include, yeast artificial chromosomes(YACs), bacterial artificial chromosomes (BACs), plant artificialchromosomes (PACs) and the like. For a general introduction to YACs,BACs, PACs and MACs as artificial chromosomes, see, e.g., Monaco andLarin (1994) Trends Biotechnol 12:280. In addition, methods for the invitro amplification of large nucleic acids linked to genetic markers arewidely available (e.g., Cheng et al. (1994) Nature 369:684, andreferences therein). Cloning systems can be created or obtained fromcommercially; see, for example, Stratagene (La Jolla, Calif.).

Vectors, Promoters and Expression Systems

The present invention includes recombinant constructs incorporating oneor more of the nucleic acid sequences described above. Such constructsinclude a vector, for example, a plasmid, a cosmid, a phage, a virus, abacterial artificial chromosome (BAC), a yeast artificial chromosome(YAC), etc., into which one or more polynucleotide sequences of interest(e.g., a marker or genetic element contributing to yield) has beeninserted, in a forward or reverse orientation. For example, the insertednucleic acid can include a chromosomal sequence or cDNA including a allor part of at least one genetic element or open reading frame (“ORF”)associated with yield. In a preferred embodiment, the construct furthercomprises regulatory sequences, including, for example, a promoter,operably linked to the sequence. Large numbers of suitable vectors andpromoters are known to those of skill in the art, and are commerciallyavailable.

As desired, the polynucleotides of the present invention, e.g., agenetic element contributing to superior agronomic performanceidentified according to the methods described herein, can be included inany one of a variety of vectors suitable for generating sense orantisense RNA, and optionally, polypeptide expression products. Suchvectors include chromosomal, nonchromosomal and synthetic DNA sequences,e.g., derivatives of SV40; bacterial plasmids; phage DNA; baculovirus;yeast plasmids; vectors derived from combinations of plasmids and phageDNA, viral DNA such as vaccinia, adenovirus, fowl pox virus,pseudorabies, adenovirus, adeno-associated virus, retroviruses and manyothers. Any vector that is capable of introducing genetic material intoa cell, and, if replication is desired, which is replicable in therelevant host can be used.

In an expression vector or expression cassette, the polynucleotidesequence of interest is physically arranged in proximity and orientationto an appropriate transcription control sequence (promoter, andoptionally, one or more enhancers) to direct mRNA synthesis. That is,the polynucleotide sequence of interest is “operably linked” to anappropriate transcription control sequence. Examples of such promotersinclude; LTR or SV40 promoter, E. coli lac or tip promoter, phage lambdaP_(L) promoter, and other promoters known to control expression of genesin prokaryotic or eukaryotic cells or their viruses. The expressionvector also contains a ribosome binding site for translation initiation,and a transcription terminator. The vector optionally includesappropriate sequences for amplifying expression. In addition, theexpression vectors optionally comprise one or more selectable markergenes to provide a phenotypic trait for selection of transformed hostcells, such as dihydrofolate reductase or neomycin resistance foreukaryotic cell culture, or such as tetracycline or ampicillinresistance in E. coli.

Additional Expression Elements

Where translation of polypeptide encoded by a nucleic acid comprising apolynucleotide sequence of the invention is desired, additionaltranslation specific initiation signals can improve the efficiency oftranslation. These signals can include, e.g., an ATG initiation codonand adjacent sequences. In some eases, for example, full-length cDNAmolecules or chromosomal segments including a coding sequenceincorporating, e.g., a QTL or an ORF associated with a QTL or QTLmarker, a translation initiation codon and associated sequence elementsare inserted into the appropriate expression vector simultaneously withthe polynucleotide sequence of interest. In such cases, additionaltranslational control signals frequently are not required. However, incases where only a polypeptide coding sequence, or a portion thereof, isinserted, exogenous translational control signals, including an ATGinitiation codon must be provided. Furthermore, the initiation codonmust be in the correct reading frame to ensure transcription of thepolynucleotide sequence of interest. Exogenous transcriptional elementsand initiation codons can be of various origins, both natural andsynthetic. The efficiency of expression can be enhanced by the inclusionof enhancers appropriate to the cell system in use (Scharf D et al.(1994) Results Probl Cell Differ 20:125-62; Bittner et al. (1987)Methods in Enzymol 153:516-544).

Generation of Transgenic Plants and Cells

The present invention also relates to host cells and organisms which aretransformed with nucleic acids corresponding to genetic elementscontributing to superior agronomic performance and other genesidentified according to the methods of the invention. For example, suchnucleic acids include chromosome segments, ORFs, and/or cDNAs orcorresponding to a sequence or subsequence included within theidentified chromosome segment or ORF. Additionally, the inventionprovides for the production of polypeptides corresponding to suchgenetic elements by recombinant nucleic acid (and expression)techniques. Host cells are genetically engineered (i.e., transduced,transfected or transformed) with the vectors of this invention (i.e.,vectors incorporating genetic elements contributing to increased yield,or other nucleic acids identified according to the methods of theinvention and as described above) which are, for example, a cloningvector or an expression vector. Such vectors include, in addition tothose described above, e.g., an agrobacterium, a virus (such as a plantvirus), a naked polynucleotide, or a conjugated polynucleotide. Thevectors are introduced into plant tissues, cultured plant cells or plantprotoplasts by a variety of standard methods including electroporation(From et al. (1985) Proc. Natl. Acad. Sci, USA 82; 5824), infection byviral vectors such as cauliflower mosaic virus (CaMV) (Hohn et al.(1982) Molecular Biology of Plant Tumors (Academic Press, New York, pp.549-560; Howell U.S. Pat. No. 4,407,956), high velocity ballisticpenetration by small particles with the nucleic acid either within thematrix of small beads or particles, or on the surface (Klein et al.(1987) Nature 327; 70), use of pollen as vector (WO 85/01856), or use ofAgrobacterium tumefaciens or A. rhizogenes carrying a T-DNA plasmid inwhich DNA fragments are cloned. The T-DNA plasmid is transmitted toplant cells upon infection by Agrobacterium tumefaciens, and a portionis stably integrated into the plant genome (Borsch et al. (1984) Science233; 496; Fraley et al. (1983) Proc. Natl. Acad. Sci. USA 80; 4803). Themethod of introducing a nucleic acid of the present invention into ahost cell is not critical to the instant invention. Thus, any method,e.g., including but not limited to the above examples, which providesfor effective introduction of a nucleic acid into a cell or protoplastcan be employed.

The engineered host cells can be cultured in conventional nutrient mediamodified as appropriate for such activities as, for example, activatingpromoters or selecting transformants. These cells can optionally becultured into transgenic plants. Plant regeneration from culturedprotoplasts is described in Evans et al. (1983) “Protoplast Isolationand Culture,” Handbook of Plant Cell Cultures 1, 124-176 (MacMillanPublishing Co., New York; Davey (1983) “Recent Developments in theCulture and Regeneration of Plant Protoplasts,” Protoplasts, pp. 12-29,(Birkhauser, Basel); Dale (1983) “Protoplast Culture and PlantRegeneration of Cereals and Other Recalcitrant Crops,” Protoplasts pp.31-41, (Birkhauser, Basel); Binding (1985) “Regeneration of Plants,”Plant Protoplasts, pp. 21-73, (CRC Press, Boca Raton).

The present invention also relates to the production of transgenicorganisms, which may be bacteria, yeast, fungi, or plants, transducedwith the nucleic acids, e.g., cloned QTL of the invention. A thoroughdiscussion of techniques relevant to bacteria, unicellular eukaryotesand cell culture may be found in references enumerated above and arebriefly outlined as follows. Several well-known methods of introducingtarget nucleic acids into bacterial cells are available, any of whichmay be used in the present invention. These include: fusion of therecipient cells with bacterial protoplasts containing the DNA, treatmentof the cells with liposomes containing the DNA, electroporation,projectile bombardment (biolistics), carbon fiber delivery, andinfection with viral vectors (discussed further, below), etc. Bacterialcells can be used to amplify the number of plasmids containing DNAconstructs of this invention. The bacteria are grown to log phase andthe plasmids within the bacteria can be isolated by a variety of methodsknown in the art (see, for instance, Sambrook). In addition, numerouskits are commercially available and can be employed according to themanufacturers instructions for the purification of plasmids frombacteria (and other cells). For their proper use, follow themanufacturer's instructions (see, for example, EasyPrep™, FlexiPrep™,both from Pharmacia Biotech; StrataClean™, from Stratagene; and,QIAprep™ from Qiagen). The isolated and purified plasmids are thenfurther manipulated to produce other plasmids, used to transfect plantcells or incorporated into Agrobacterium tumefaciens related vectors toinfect plants. Typical vectors contain transcription and translationterminators, transcription and translation initiation sequences, andpromoters useful for regulation of the expression of the particulartarget nucleic acid. The vectors optionally comprise generic expressioncassettes containing at least one independent terminator sequence,sequences permitting replication of the cassette in eukaryotes, orprokaryotes, or both, (e.g., shuttle vectors) and selection markers forboth prokaryotic and eukaryotic systems. Vectors are suitable forreplication and integration in prokaryotes, eukaryotes, or preferablyboth. See, Gillman & Smith (1979) Gene 8:81; Roberts et al. (1987)Nature 328:731; Schneider et al. (1995) Protein Expr. Purif 6435; 10;Ausubel, Sambrook, Berger (all supra). A catalogue of Bacteria andBacteriophages useful for cloning is provided, e.g., by the ATCC, e.g.,The ATCC Catalogue of Bacteria and Bacteriophage (1992) Gherna et al.(eds) published by the ATCC. Additional basic procedures for sequencing,cloning and other aspects of molecular biology and underlyingtheoretical considerations are also found in Watson at al. (1992).Recombinant DNA, Second Edition, Scientific American Books, NY.

Transforming Nucleic Acids into Plants.

Embodiments of the present invention pertain to the production oftransgenic plants comprising the cloned nucleic acids, e.g., chromosomesegments, isolated ORFs, and cDNAs associated with genetic elementsidentified by their proximity to the markers of the invention.Techniques for transforming plant cells with nucleic acids are generallyavailable and can be adapted to the invention by the use of nucleicacids encoding or corresponding to chromosome segments, subsequences,e.g., ORFs, and the like. In addition to Berger, Ausubel and Sambrook(infra), useful general references for plant cell cloning, culture andregeneration include Jones (ed) (1995) Plant Gene Transfer andExpression Protocols—Methods in Molecular Biology, Volume 49 HumanaPress Towata N.J.; Payne et al. (1992) Plant Cell and Tissue Culture inLiquid Systems John Wiley & Sons, Inc. New York, N.Y. (Payne); andGamborg and Phillips (eds) (1995) Plant Cell, Tissue and Organ Culture;Fundamental Methods Springer Lab Manual, Springer-Verlag (BerlinHeidelberg New York) (Gamborg). A variety of cell culture media aredescribed in Atlas and Parks (eds) The Handbook of Microbiological Media(1993) CRC Press, Boca Raton, Fla. (Atlas). Additional information forplant cell culture is found in available commercial literature such asthe Life Science Research Cell Culture Catalogue (1998) fromSigma-Aldrich, Inc (St Louis, Mo.) (Sigma-LSRCCC) and, e.g., the PlantCulture Catalogue and supplement (1997) also from Sigma-Aldrich, Inc (StLouis, Mo.) (Sigma-PCCS). Additional details regarding plant cellculture are found in Croy, (ed.) (1993) Plant Molecular Biology BiosScientific Publishers, Oxford, U.K.

The nucleic acid constructs of the invention, e.g., plasmids, cosmids,artificial chromosomes, DNA and RNA polynucleotides, are introduced intoplant cells, either in culture or in the organs of a plant by a varietyof conventional techniques. Where the sequence is expressed, thesequence is optionally combined with transcriptional and translationalinitiation regulatory sequences which direct the transcription ortranslation of the sequence from the exogenous DNA in the intendedtissues of the transformed plant.

Isolated nucleic acids can be introduced into plants according to any ofa variety of techniques known in the art. Techniques for transforming awide variety of higher plant species are well known and described in thetechnical, scientific, and patent literature. See, for example, Weisinget al. (1988) Ann. Rev. Genet. 22:421-477.

For example plasmids, cosmids, phage, naked or variously conjugated-DNApolynucleotides, (e.g., polylysine-conjugated DNA, peptide-conjugatedDNA, liposome-conjugated DNA, etc.), or artificial chromosomes, can beintroduced directly into the genomic DNA of the plant cell usingtechniques such as electroporation and microinjection of plant cellprotoplasts, or the DNA constructs can be introduced directly to plantcells using ballistic methods, such as DNA particle bombardment.

Microinjection techniques for injecting e.g., cells, embryos, callus andprotoplasts, are known in the art and well described in the scientificand, patent literature. For example, a number of methods are describedin Jones (ed) (1995) Plant Gene Transfer and ExpressionProtocols—Methods in Molecular Biology, Volume 49 Humana Press TowataN.J., as well as in the other references noted herein and available inthe literature.

For example, the introduction of DNA constructs using polyethyleneglycol precipitation is described in Paszkowski, et al., EMBO J. 3:2717(1984). Electroporation techniques are described in Fromm, et al., Proc.Nat'l. Acad. Sci. USA 82:5824 (1985). Ballistic transformationtechniques are described in Klein, et al., Nature 327:70-73 (1987).Additional details are found in Jones (1995) and Gamborg and Phillips(1995), supra, and in U.S. Pat. No. 5,990,387.

Alternatively, and in some cases preferably, Agrobacterium mediatedtransformation is employed to generate transgenic plants,Agrobacterium-mediated transformation techniques, including disarmingand use of binary vectors, are also well described in the scientificliterature. See, for example Horsch, et al. (1984) Science 233:496; andFraley et al. (1984) Proc. Nat'l. Acad. Sci. USA 80:4803 and recentlyreviewed in Hansen and Chilton (1998) Current Topics in Microbiology240:22 and Das (1998) Subcellular Biochemistry 29: Plant MicrobeInteractions pp 343-363.

The DNA constructs may be combined with suitable T-DNA flanking regionsand introduced into a conventional Agrobacterium tumefaciens hostvector. The virulence functions of the Agrobacterium tumefaciens hostwill direct the insertion of the construct and adjacent marker into theplant cell DNA when the cell is infected by the bacteria. See, U.S. Pat.No. 5,591,616. Although Agrobacterium is useful primarily in dicots,certain monocots can be transformed by Agrobacterium. For instance,Agrobacterium transformation of maize is described in U.S. Pat. No.5,550,318.

Other methods of transfection or transformation include (I)Agrobacterium rhizogenes-mediated transformation (see, e.g.,Lichtenstein and Fuller (1987) In: Genetic Engineering, vol. 6, PWJRigby, Ed., London, Academic Press; and Lichtenstein; C. P., and Draper(1985) In: DNA Cloning, Vol. II, D. M. Glover, Ed., Oxford, IRI Press;WO 88/02405, published Apr. 7, 1988, describes the use of A. rhizogenesstrain A4 and its Ri plasmid along with A. tumefaciens vectors pARC8 orpARC16 (2) liposome-mediated DNA uptake (see, e.g., Freeman et al,(1984) Plant Cell Physiol. 25:1353), (3) the vortexing method (see,e.g., Kindle (1990) Proc. Natl. Acad. Sci., (USA) 87:1228.

DNA can also be introduced into plants by direct DNA transfer intopollen as described by Zhou et al. (1983) Methods in Enzymology,101:433; D. Hess (1987) Intern Rev. Cytol. 107:367; Luo et al. (1988)Plant Mol. Biol. Reporter 6:165. Expression of polypeptide coding genescan be obtained by injection of the DNA into reproductive organs of aplant as described by Pena et al. (1987) Nature 325:274. DNA can also beinjected directly into the cells of immature embryos and the desiccatedembryos rehydrated as described by Neuhaus et al. (1987) Theor. Appl.Genet. 75:30; and Benbrook et al. (1986) in Proceedings Bio ExpoButterworth, Stoneham, Mass., pp. 27-54. Additionally, a variety ofplant viruses that can be employed as vectors are known in the art andinclude cauliflower mosaic virus (CaMV), geminivirus, brome mosaicvirus, and tobacco mosaic virus.

Regeneration of Transgenic Plants

Transformed plant cells which are derived by any of the abovetransformation techniques can be cultured to regenerate a whole plantwhich possesses the transformed genotype and thus the desired phenotype.Such regeneration techniques rely on manipulation of certainphytohormones in a tissue culture growth medium, typically relying on abiocide and/or herbicide marker which has been introduced together withthe desired nucleotide sequences. Plant regeneration from culturedprotoplasts is described in Evans et al. (1983) Protoplasts Isolationand Culture, Handbook of Plant Cell Culture pp. 124-176, MacmillianPublishing Company, New York; and Binding (1985) Regeneration of Plants,Plant Protoplasts pp. 21-73, CRC Press, Boca Raton. Regeneration canalso be obtained from plant callus, explants, somatic embryos (Dandekaret al. (1989) J. Tissue Cult. Meth. 12:145; McGranahan, et al. (1990)Plant Cell Rep. 8:512) organs, or parts thereof. Such regenerationtechniques are described generally in Klee et al. (1987)., Ann. Rev. ofPlant Phys. 38:467-486. Additional details are found in Payne (1992) andJones (1995), both supra, and Weissbach and Weissbach, eds. (1988)Methods for Plant Molecular Biology Academic Press, Inc., San Diego,Calif. This regeneration and growth process includes the steps ofselection of transformant cells and shoots, rooting the transformantshoots and growth of the plantlets in soil. These methods are adapted tothe invention to produce trans genic plants bearing QTLs and other genesisolated according to the methods of the invention.

In addition, the regeneration of plants containing the polynucleotide ofthe present invention and introduced by Agrobacterium into cells of leafexplants can be achieved as described by Horsch et al. (1985) Science227:1229-1231. In this procedure, transformants are grown in thepresence of a selection agent and in a medium that induces theregeneration of shoots in the plant species being transformed asdescribed by Fraley et al. (1983) Proc. Natl. Acad. Sci. (U.S.A.)80:4803. This procedure typically produces shoots within two to fourweeks and these transformant shoots are then transferred to anappropriate root-inducing medium containing the selective agent and anantibiotic to prevent bacterial growth. Transgenic plants of the presentinvention may be fertile or sterile.

In construction of recombinant expression cassettes of the invention,which include, for example, an ORF associated with a marker or geneticelement contributing to yield, a plant promoter fragment is optionallyemployed which directs expression of a nucleic acid in any or alltissues of a regenerated plant. Examples of constitutive promotersinclude the cauliflower mosaic virus (CaMV) 35S transcription initiationregion, the or 2′-promoter derived from T-DNA of Agrobacteriumtumefaciens, and other transcription initiation regions from variousplant genes known to those of skill. Alternatively, the plant promotermay direct expression of the polynucleotide of the invention in aspecific tissue (tissue-specific promoters) or may be otherwise undermore precise environmental control (inducible promoters). Examples oftissue-specific promoters under developmental control include promotersthat initiate transcription only in certain tissues, such as fruit,seeds, or flowers.

Any of a number of promoters which direct transcription in plant cellscan be suitable. The promoter can be either constitutive or inducible.In addition to the promoters noted above, promoters of bacterial originwhich operate in plants include the octopine synthase promoter, thenopaline synthase promoter and other promoters derived from native Tiplasmids. See, Herrara-Estrella et al., (1983), Nature, 303:209. Viralpromoters include the 35S and 19S RNA promoters of cauliflower mosaicvirus. See, Odell et at (1985) Nature, 313:810. Other plant promotersinclude the ribulose-1,3-bisphosphate carboxylase small subunit promoterand the phaseolin promoter. The promoter sequence from the E8 gene andother genes may also be used. The isolation and sequence of the E8promoter is described in detail in Deikman and Fischer (1988) EMBO J.7:3315. Many other promoters are in current use and can be coupled to anexogenous DNA sequence to direct expression of the nucleic acid.

If expression of a polypeptide, including those encoded by QTL or othernucleic acid, is desired, a polyadenylation region at the 3′-end of thecoding region is typically included. The polyadenylation region can bederived from the natural gene, from a variety of other plant genes, orfrom, e.g., T-DNA.

The vector comprising the sequences (e.g., promoters or coding regions)from genes encoding expression products and transgenes of the inventionwill typically include a nucleic acid subsequence, a marker gene whichconfers a selectable, or alternatively, a screenable, phenotype on plantcells. For example, the marker may encode biocide tolerance,particularly antibiotic tolerance, such as tolerance to kanamycin, G418,bleomycin, hygromycin, or herbicide tolerance, such as tolerance tochlorosluforon, or phosphinothricin (the active ingredient in theherbicides bialaphos or Basta). See, e.g., Padgette et al. (1996) In:Herbicide-Resistant Crops (Duke, ed.), pp 53-84, CRC Lewis Publishers,Boca Raton (“Padgette, 1996”). For example, crop selectivity to specificherbicides can be conferred by engineering genes into crops which encodeappropriate herbicide metabolizing enzymes from other organisms, such asmicrobes. See, Vasil (1996) in: Herbicide-Resistant Crops (Duke, ed.),pp 85-91, CRC Lewis Publishers, Boca Raton) (“Vasil”, 1996).

One of skill will recognize that after the recombinant expressioncassette is stably incorporated in transgenic plants and confirmed to beoperable, it can be introduced into other plants by sexual crossing. Anyof a number of standard breeding techniques can be used, depending uponthe species to be crossed. In vegetatively propagated crops, maturetransgenic plants can be propagated by the taking of cuttings or bytissue culture techniques to produce multiple identical plants.Selection of desirable transgenics is made and new varieties areobtained and propagated vegetatively for commercial use. In seedpropagated crops, mature transgenic plants can be self crossed toproduce a homozygous inbred plant. The inbred plant produces seedcontaining the newly introduced heterologous nucleic acid. These seedscan be grown to produce plants that would produce the selectedphenotype. Parts obtained from the regenerated plant, such as flowers,seeds, leaves, branches, fruit, and the like are included in theinvention, provided that these parts comprise cells comprising theisolated nucleic acid of the present invention. Progeny and variants,and mutants of the regenerated plants are also included within the scopeof the invention, provided that these parts comprise the introducednucleic acid sequences.

Transgenic plants expressing a polynucleotide of the present inventioncan be screened for transmission of the nucleic acid of the presentinvention by, for example, standard immunoblot and DNA detectiontechniques. Expression at the RNA level can be determined initially toidentify and quantitate expression-positive plants. Standard techniquesfor RNA analysis can be employed and include PCR amplification assaysusing oligonucleotide primers designed to amplify only the heterologousRNA templates and solution hybridization assays using heterologousnucleic acid-specific probes. The RNA-positive plants can then analyzedfor protein expression by Western immunoblot analysis using thespecifically reactive antibodies of the present invention. In addition,in situ hybridization and immunocytochemistry according to standardprotocols can be done using heterologous nucleic acid specificpolynucleotide probes and antibodies, respectively, to localize sites ofexpression within transgenic tissue. Generally, a number of trans genielines are usually screened for the incorporated nucleic acid to identifyand select plants with the most appropriate expression profiles.

A preferred embodiment is a transgenic plant that is homozygous for theadded heterologous nucleic acid; i.e., a transgenic plant that containstwo added nucleic acid sequences, one gene at the same locus on eachchromosome of a chromosome pair. A homozygous transgenic plant can beobtained by sexually mating (selfing) a heterozygous transgenic plantthat contains a single added heterologous nucleic acid, germinating someof the seed produced and analyzing the resulting plants produced foraltered expression of a polynucleotide of the present invention relativeto a control plant (i.e., native, non-transgenic). Back-crossing to aparental plant and out-crossing with a non-transgenic plant are alsocontemplated.

General Molecular Biology References

In the context of the invention, e.g., identifying, monitoring and/orcloning molecular markers and/or other loci, nucleic acids and/orproteins are manipulated according to well known molecular biologytechniques. Detailed protocols for numerous such procedures aredescribed in, e.g., in Ausubel et al. Current Protocols in MolecularBiology (supplemented through 2000) John Wiley & Sons, New York(“Ausubel”); Sambrook et al. Molecular Cloning—A Laboratory Manual (2ndEd.), Vol. 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.,1989 (“Sambrook”), and Berger and Kimmel Guide to Molecular CloningTechniques, Methods in Enzymology volume 152 Academic Press, Inc., SanDiego, Calif. (“Berger”).

In addition to the above references, protocols for in vitroamplification techniques, such as the polymerase chain reaction (PCR),the ligase chain reaction (LCR), Qβ-replicase amplification, and otherRNA polymerase mediated techniques (e.g., NASBA), useful e.g., foramplifying cDNA probes of the invention, are found in Mullis et al.(1987) U.S. Pat. No. 4,683,202; PCR Protocols A Guide to Methods andApplications (Innis et al. eds) Academic Press Inc. San Diego, Calif.(1990) (“Innis”); Arnheim and Levinson (1990) C&EN 36; The Journal OfNIH Research (1991) 3:81; Kwoh et al. (1989) Proc Natl Acad Sci USA 86,1173; Guatelli et al. (1990) Proc Nail Acad Sci USA 87:1874; Lomell etal. (1989) J Clin Chem 35:1826; Landegren et at (1988) Science 241:1077;Van Brunt (1990) Biotechnology 8:291; Wu and Wallace (1989) Gene 4: 560;Barringer et al. (1990) Gene 89:117, and Sooknanan and Malek (1995)Biotechnology 13:561 Additional methods, useful for cloning nucleicacids in the context of the present invention, include Wallace et al.U.S. Pat. No. 5,426,039. Improved methods of amplifying large nucleicacids by PCR are summarized in Cheng et al. (1994) Nature 369:684 andthe references therein.

Certain polynucleotides of the invention, e.g., oligonucleotides can besynthesized utilizing various solid-phase strategies involvingmononucleotide- and/or trinucleotide-based phosphoramidite couplingchemistry. For example, nucleic acid sequences can be synthesized by thesequential addition of activated monomers and/or timers to an elongatingpolynucleotide chain. See e.g., Caruthers, M. H. et al. (1992) MethEnzymol 211:3.

In lieu of synthesizing the desired sequences, essentially any nucleicacid can be custom ordered from any of a variety of commercial sources,such as The Midland Certified Reagent Company (mcrc@oligos.com), TheGreat American Gene Company (www.genco.com), ExpressGen, Inc.(www.expressgen.com), Operon Technologies, Inc. (www.operon.com), andmany others.

Similarly, commercial sources for nucleic acid and protein microarraysare available, and include, e.g., Affymetrix, Santa Clara, Calif.(http://www.affymetrix.com/); and Incyte, Palo Alto, Calif. (on theworld wide web at incyte.com); and Ciphergen Biosciences, Fremont,Calif. (at ciphergen.com).

High Throughput Screening

In one aspect of the invention, the determination of genetic markeralleles is performed by high throughput screening. High throughputscreening involves providing a library of genetic markers, e.g., SSRprimers, ASH primers and probes, RFLPs, AFLPs, isozymes, specificalleles and variable sequences, including SSR, RAPD and the like. Suchlibraries are then screened against plant genomes to generate a“fingerprint” for each plant under consideration. In some cases apartial fingerprint comprising a sub-portion of the markers is generatedin an area of interest. Once the genetic marker alleles of a plant havebeen identified, the correspondence between one or several of the markeralleles and a desired phenotypic trait is determined through statisticalassociations based on the methods of this invention.

High throughput screening can be performed in many different formats.Hybridization can take place in a 96-, 324-, or a 1524-well format or ina matrix on a silicon chip or other format.

A number of well-known robotic systems have been developed for highthroughput screening, particularly in a 96 well format. These systemsinclude automated work stations like the automated synthesis apparatusdeveloped by Takeda Chemical Industries, LTD. (Osaka, Japan) and manyrobotic systems utilizing robotic arms (Zymate II, Zymark Corporation,Hopkinton, Mass.; ORCA™, Beckman Coulter, Fullerton Calif.). Any of theabove devices are suitable for use with the present invention. Thenature and implementation of modifications to these devices (if any) sothat they can operate as discussed herein will be apparent to personsskilled in the relevant art.

In addition, high throughput screening systems themselves arecommercially available (see, e.g., Zymark Corp., Hopkinton, Mass.; AirTechnical Industries, Mentor, Ohio; Beckman Instruments, Inc. Fullerton,Calif.; Precision Systems, Inc., Natick, Mass., etc.). These systemstypically automate entire procedures including all sample and reagentpipetting, liquid dispensing, timed incubations, and final readings ofthe microplate or membrane in detector(s) appropriate for the assay.These configurable systems provide high throughput and rapid start up aswell as a high degree of flexibility and customization. Themanufacturers of such systems provide detailed protocols for the use oftheir products in high throughput applications.

In one variation of the invention, solid phase arrays are adapted forthe rapid and specific detection of multiple polymorphic nucleotides.Typically, a nucleic acid probe is linked to a solid support and atarget nucleic acid is hybridized to the probe. Either the probe, or thetarget, or both, can be labeled, typically with a fluorophore. If thetarget is labeled, hybridization is evaluated by detecting boundfluorescence. If the probe is labeled, hybridization is typicallydetected by quenching of the label by the bound nucleic acid. If boththe probe and the target are labeled, detection of hybridization istypically performed by monitoring a color shift resulting from proximityof the two bound labels.

In one embodiment, an array of probes are synthesized on a solidsupport. Using chip masking technologies and photoprotective chemistry,it is possible to generate ordered arrays of nucleic acid probes. Thesearrays, which are known, e.g., as “DNA chips” or as very large scaleimmobilized polymer arrays (VLSIPS™ arrays) can include millions ofdefined probe regions on a substrate having an area of about 1 cm² toseveral cm².

In another embodiment, capillary electrophoresis is used to analyzepolymorphism. This technique works best when the polymorphism is basedon size, for example, SSR and AFLP. This technique is described indetail in U.S. Pat. Nos. 5,534,123 and 5,728,282. Briefly, capillaryelectrophoresis tubes are filled with the separation matrix. Theseparation matrix contains hydroxyethyl cellulose, urea and optionallyfor amide. The SSR and AFLP samples are loaded onto the capillary tubeand electrophoresed. Because of the small amount of sample andseparation matrix required by capillary electrophoresis, the run timesare very short. The molecular sizes and therefore, the number ofnucleotides present in the nucleic acid sample is determined bytechniques described herein. In a high throughput format, many capillarytubes are placed in a capillary electrophoresis apparatus. The samplesare loaded onto the tubes and electrophoresis of the samples is runsimultaneously. See, Mathies and Huang, (1992) Nature 359:167.

EXAMPLES

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims.

Example 1 Summary of Breeding Bias Data-Favorable Allelic Forms byGeographic Region

Tables 3 through 9 enumerate favorable allelic forms of chromosomesegments contributing to superior agronomic performance in differentgeographic regions and growing environments. * 95% significance level.** 99% significance level. UM indicates an unmapped locus. Detailsregarding SSR and ASH markers and alleles are provided in Appendices Ithrough IV. The geographic regions are defined with respect to thefollowing reference points.

Central Region: Central Midwest United States centered around DesMoines, Iowa.

Canada Region Eastern Canada centered around Chatham, Ontario.

North Region Northern Midwest United States centered around RedwoodFalls, Minn.

Illinois Region Northern and central Illinois centered around Champaign,Ill.

East Region: Eastern Midwest United States centered around Napoleon,Ohio.

Mid South Region: Mid Southern United States centered around Hamil, Ill.

South Region Southern United States centered around Memphis, Tenn.

Example 2 Favorable Soybean Alleles and “Target Genotype(s)” forAdaptation to Environments Similar to Those Found in Iowa USA (CentralRegion)

Three separate breeding bias analyses were conducted to identifyfavorable alleles that have provided adaptation to geographic regionsthat are typical of Iowa, USA. The first analysis used 41 elite lines asthe “elite population” adapted to Iowa, the second analysis included alarger sample of 71 elite lines (Table 1, Central Region) and the thirdanalysis, performed following the 2003 growing season, used 86 elitelines (Table 1, Central Region 2003). In all cases, the elite populationwas chosen as a representative sample of elite lines that yield well inIowa and/or are good parents for developing elite lines adapted to Iowa.Each successive analysis was run after more elite line marker data wasavailable and was therefore considered more rigorous. Despite this fact,the results of the first analysis were very similar and demonstratedthat the breeding bias method is useful even with smaller datasets.Herein, results of the second (A) and third (13) analyses are reporteddue to the larger sample size of the elite population.

A computer program described in U.S. Pat. No. 5,437,697 that simulatesthe flow of alleles from ancestors to elite lines at each marker locuswas used to determine what percent of the elite lines would be expectedto inherit each marker allele by chance alone. By doing multipleiterations of the simulation process (10,000 iterations in this case), astatistical measure of random result variation was obtained. The averagefrequency of each allele in the simulated elite population (from the10,000 iterations) is herein referred to as the “expected frequency”(EXP) of each marker allele. The expected frequency was then compared tothe “observed frequency” (OBS) which is simply a count of how many lineswithin the elite population actually contain a given allele divided bythe total number of elite lines examined. By comparing the observedresults to the results of the simulation process, one can determine howoften the observed results would be expected to occur by chance alone.If the observed results would only be expected 5% of the time or lessdue to chance (i.e. a LOD score of 1.3 or greater), it is safe to assumethat the observed results did not occur by chance alone. In this case,selection for grain yield must have biased selection towards one or morealleles at the locus in question. The allele(s) that occurred at higherfrequency than expected were therefore labeled as “favorable alleles”and those occurring at a lower frequency than expected were labeled as“unfavorable alleles.” Favorable alleles are those that must havecontributed either directly or indirectly to higher grain yield.

The statistically significant results of the Iowa (A) analysis are shownin Table 10. Since only the major ancestors were genotyped in theanalysis, occasionally an allele that was detected in the elitepopulation was not always found in the ancestral population. Thisresulted in an unusually high LOD score since the frequency of theallele in the ancestral population was assumed to be zero. It is alsoreasonable to assume that a favorable allele should have increased infrequency by some threshold percentage before that allele should beconsidered generally “favorable” over the wide range of environmentsencountered by soybean breeders over the past century. For thesereasons, an allele with a LOD score of greater than or equal to 1.3 (95%confidence) AND an observed frequency of at least 25% higher thanexpected was considered to be “favorable” and “significant” from both apractical and statistical view. Alleles with these criteria plus a LODscore of 2.0 or greater (99% confidence), were considered to be“favorable” and “highly significant.” Alleles with a LOD scores ofgreater than 1.3 but that did not increase in frequency by at least 25%over the past century were considered statistically significant but notenough to be considered generally favorable over most environments.

The term “LOD” score refers to the negative inverse log(base 10)probability that the observed frequency of an allele in the elitepopulation could be attributed to chance alone. More specifically, theLOD score is a measure of the number of rounds that the breeding biassimulation generated an allele frequency at least as extreme as thatobserved in the actual elite population of soybean lines. The formulafor a LOD score of a “favorable” allele is: −1.0×log 10 (f) where:f=(the number of rounds of simulation where the observed allelefrequency in the elite population was greater than that generated by thesimulated allele frequency) divided by (the total number of rounds ofsimulation).

For example, a LOD score of >1.30 is analogous to a probability of <0.05that the observed allele frequency in the elite population was due tochance alone; a LOD score of >2.00 is analogous to a probability of<0.01 that the observed allele frequency in the elite population was dueto chance alone; a LOD score of >3.00 is analogous to a probability of<0.001 that the observed allele frequency in the elite population wasdue to chance alone; a LOD score of 4.00 is analogous to a probabilityof 0.0001 that the observed allele frequency in the elite population wasdue to chance alone. Since only 10,000 rounds of simulation wereconducted, the maximum LOD score observed did not go any higher than4.00.

Based on the Iowa analysis, out of 1540 alleles over 309 genetic markerloci, a total of 94 alleles showed evidence of being favorable by theaforementioned definition: a LOD score of at least 1.30 (95% confidencelevel) and an increase in allele frequency of at least 25% more thanexpected by random inheritance. Since some of these favorable allelesare closely linked (i.e., less than 10 CM apart according to independentmapping studies), not all 94 alleles are diagnostic of unique QTL loci.Therefore, the markers were divided into genomic regions with theassumption that markers on the same chromosome that are approximately 10CM or greater apart are probably diagnostic of different QTL loci.

In addition to listing the favorable alleles that span the genome, Table10 indicates the favorable allele with the best statistical score and/orthe most marker data within each predefined genomic region. The “best”marker allele to use for each region can also be chosen based on whichmarker works best in the laboratory. In most cases, the marker with thehighest LOD score and highest % difference of expected and observedallele frequency was considered to “best” marker in its genomic region.For example, on chromosome C1, there are 4 markers that map to positions95.8 through 99.0-Satt399, Satt361, P10639A-1, and Satt190. These areall within 10 CM of each other and therefore were assigned to the samegenomic region #29. Among these 4 markers, markers Satt399 and Satt190had the highest possible LOD score of 4.0 when 10,000 iterations of asimulation were done. This means that even after 10,000 iterations weredone, there was not even 1 iteration where the simulation producedresults more extreme than that observed in the actual elite population.A LOD score of 4.0 is merely the inverse log of the probability (1 in10,000) that the results obtained at these loci was due to chance alone.From there, one can decide which of these 2 loci would be the bestmarker to identify the favorable allele in that region. In this case,Satt190 was chosen as the best marker in that region because the resultswere based on data from 65 as opposed to 61 elite lines. Although atotal of 71 elite lines were genotyped for this marker, 6 elite lineshad missing data for this locus.

Once a desirable marker is identified and the favorable allele of thatmarker is determined, selection for that favorable allele in a lab assaycan then be used for MAS to identify plants that have the favorableallele. The MAS process can be done at multiple loci simultaneously toselect for plants that contain the maximum number of favorable allelesthat span the genome. This genome-wide group of favorable alleles uponwhich selection is based is herein referred to as the “Target Genotype.”Table 11 shows the genome-wide Target Genotype for the best locus fromeach genomic region that meets the criteria of “significant”—i.e. LODscore of at least 1.3 and an increase in allele frequency of at least25% greater than expected by chance alone. A total of 57 favorablealleles, each representing what was favored by selection in a differentgenomic region, fit these criteria to make the Target Genotype at LOD1.3(Table 11, *). If one raises the statistical cutoff to a LOD of 2.0, theTarget Genotype focuses in on 30 favorable alleles (Table 11, **). Forpurposes of MAS, one would want to select for the favorable alleles withthe highest statistical significance first. Hence, the logical path forbreeding purposes would be to select among segregating progeny for the30 locus Target Genotype first. Once these loci are fixed for thefavorable alleles, the other 27 loci in the 57-locus Target Genotypecould be the focus of selection. If resources are unlimited, one couldconceivably work with all 57 loci. Loci can also be weighted based ontheir statistical significance for selection purposes.

Following the 2003 growing season, an additional analysis (B) wasperformed increasing the number of elite parental lines from 71 to 86,and evaluating an additional 265 SSR markers. The elite lines used inthis analysis are given in Table 1.

Significant favorable alleles are given in Table 12. Because more elitelines were used to define the population, and because some previouslyabsent data was obtained for the previously employed markers, somedifferences in the statistical LOD scores were observed for the markersin Analysis B as compared to prior Analysis A. Almost all of thepreviously identified markers were confirmed in the expanded analysis,and several new markers in the same genomic regions were found to havehigher LOD scores. Accordingly, these new markers are also deemed to beuseful in defining the target genotype and identifying and trackingfavorable alleles in soybean germplasm.

The Target Genotype is actually a consensus marker genotype that theelite population has been moving towards as the result of selection.Since this genotype is now defined by specific markers and specificfavorable alleles, it is possible to practice selection by genotypeinstead of the inefficient and slow process of selection based onphenotype. The resolution of the consensus genotype is limited only bythe genomic coverage provided by the genetic markers that are availablefor MAS.

Example 3 Status of Favorable Alleles in Soybean Variety A3127

The following example is given to illustrate how the favorable allelesidentified in the previous example came together in the most famoustransgressive segregant in soybean breeding history. Most new soybeanvarieties are a small improvement over either of their parents in termsof yield. Yield progress per cycle (5 to 6 years) of breeding iscommonly a few percent better than either parent. However, in the early1980's a variety called A3127 was developed that was much better thaneither parent (˜10% better than either parent). In fact, A3127 isprobably one of the few lines that all soybean breeders are familiarwith because it was famous for being the highest yielding variety ofit's time (early 1980's). Prior to commercialization A3127 proved to bemuch higher yielding than either of its two parents Williams and Essex.A3127 was so popular, that it became the most frequently used parent insoybean breeding history. Since A3127 is adapted to Iowa, we studied themarker profiles of Williams, Essex, and A3127 at the 30 preferred “yieldgene” loci identified for the Iowa geographic zone. We found thatWilliams and Essex differ at 23 out of 30 of these loci (Table 13). Outof the 23 segregating loci, Williams supplied 13 of the favorablealleles and Essex supplied the other 10 favorable alleles. If thesereally are the major yield loci, one would expect that A3127 would havesignificantly more favorable alleles than either parent. Amazingly,A3127 had all 23 favorable alleles. Such a segregant would only beexpected to happen by chance in 1 out of >8 million progeny (0.5²³)unless these loci really are diagnostic of yield and A3127 is truly aunique segregant. The marker genotype of A3127 is therefore consistentwith the hypothesis that these 30 marker loci are diagnostic of yield.

Example 4 Segregation for Yield in Near-Isogenic Sublines

Typically, modern soybean varieties originate from a single plantselected from a partially inbred (commonly F3) population that wasgenerated from a controlled mating between genetically differentparents. Seed of the new variety is then multiplied by subsequentpooling (or “bulking”) of seed from the self-pollinating progeny of theselected F3 plant. For any locus in the original F3 plant that washeterozygous, the resulting inbred variety will eventually become amixture of the two homozygotes at said locus. For commercial purposes,soybean varieties are purified for obvious visual traits (e.g., flowercolor, hilum color, maturity, and other visual traits that are highlyheritable and controlled by one or a few genes) but often harborresidual genetic variation that is not obvious to the naked eye. Geneticmarkers can be used, to detect loci contributing to that variation thatare still segregating within a so-called “pure line.” Since the breedingbias analysis identifies which marker loci have been affected byselection for seed yield, these markers are ideal tools to identifygenetic differences among plants within the original heterogeneousvariety that may translate into seed yield improvement. These markerscan be used to separate the original heterogeneous variety into“near-isogenic” sublines that differ at specific genetic loci.

For example, samples of seed from individual self-pollinated progenyselected from the variety are genotyped, and seed sharing a commonallele at one or more identified marker loci is pooled to produce asubline. Such sublines are genetically distinct from one another. In“blind” sublining (i.e., sublining unassisted by marker data) there isno guarantee that the generated sublines are genetically distinct. Bypooling seed of many plants with a homogenous marker genotype, enoughseed for controlled replicate trials can be obtained in one generation.This is preferable to blind sublining in several ways. First, the onlyway to attempt genetic homogeneity without markers is to pool the seedof a single plant that may or may not be genetically distinct from theseed of other single plants. Second, a single plant can only supplyenough seed for a short row yield test in one environment. Therefore,blind sublining requires subsequent generations of seed increase toobtain enough seed for highly replicated yield trials that are necessaryfor reliable yield comparisons. Third, even if phenotypic differencesare observed with blind sublining, no genomic information is gained forfuture use. By comparing the field performance of such marker-basedsublines in controlled experiments, one can determine the phenotypiceffect (e.g., with respect to yield) of each allele (and thecorresponding genomic region, if the marker is mapped) in a givengenetic background. This is particularly useful for traits, such asyield, in which gene-by-gene and gene-by-environment interactions play asubstantial role in phenotype. If one subline performs significantlybetter than the other, the better subline can be multiplied and releasedas an improved version of the original variety.

Because selection that is based on genotype (e.g., qualitative DNApolymorphism) and then confirmed with a phenotypic difference is morereliable and heritable than selection based on phenotype alone (i.e.,blind sublining), the improvement in phenotype in a blind subline isless likely to be heritable, and unlikely to be repeated in subsequentgenerations. In contrast, marker-based sublining not only providesuseful genomic information, but it also improves the heritability andreproducibility of selected traits. Thus, marker-based “subliming” canbe a powerful tool for both product development and to determine thephenotypic effect of individual loci in a given genetic background.

In accordance with this method, the genetic markers identified throughbreeding bias were shown to be effective tools to select within elitelines for residual yield gain. When genotyping elite lines with geneticmarkers, 8 random plants from each elite line are routinely sampled andbulked. If the elite line is a 50:50 mixture of two homozygotes at agiven locus, a random 8-plant sample will detect both alleles >99% ofthe time. Using this sampling procedure, segregation within commercialsoybean lines is detected at an average of about 4% of the marker lociassayed (e.g., when assaying the “best” marker loci for each chromosomalregion).

In the following exemplary trial, six elite lines were examined. Two ofthe elite lines (91B91 and 92M70) were shown to be segregating at twomarker loci each while the other 4 elite lines (92B05, 93B01, 93M80, and93M90) were segregating at one marker locus as indicated in Table 14.

To develop the sublines, leaf tissue from individual plants of each ofthe above elite lines was genotyped with the marker(s) segregating inthe originating line. Progeny seed from individual homozygous plants ofthe same marker genotype were then pooled to obtain enough seed of eachsubline to conduct replicated yield trials. The number of plant used tocreate each subline is shown in Table 15.

To determine the relative seed yield, sublines derived from the sameelite line were planted in a split plot field design at between 5 and 14locations, treating each location as an individual replication.Locations were chosen to span the soybean growing region of appropriatematurity zone for the lines being tested in the Midwestern UnitedStates. Sublines derived from a given elite line were randomly assignedto split plots within each main plot. Each split plot consisted of two12-foot long rows of a given sublime that were spaced 30 inches apart.Seed yield was measured at maturity and converted to bushels per acre (1bushel-60 pounds).

Significant yield differences between isolines were detected in 3 of the6 isoline tests. Since the isolines tested above were derived frompooling many plants of similar genotype, one can reasonably assume thatthe possibility of residual segregation at other independent loci wasrandomized and not the source of the yield difference between isolines.The magnitude of the significant yield differences (5.4 to 6.2% betweensublines or 2.7 to 3.1% better than the original mixed variety) is ofsimilar magnitude as yield improvements that can typically be obtainedusing much more exhaustive breeding efforts. New soybean varietiesdeveloped without the aid of yield gene markers can easily requirehundreds to thousands of yield plots to identify a new variety that is 2or 3% better than it's best parent. This method can be used to identifyyield gains of similar magnitude with very limited resources (2isolines×14 replications=28 plots per test). In addition, by basingselection on a real genetic difference at a locus showing historicalbreeding bias, the confidence that the yield differences detected aregenetically based (as opposed to environmental or experimental error) issubstantially increased.

Additionally, these results confirm that the effects of epistasis,gene-by-environment interactions and/or recombination between the markerallele identified by breeding bias and the genetic element underlyingyield improvements, while prevalent, do not impair selection of improvedsoybean varieties, especially if care is taken to identify residualvariation and select appropriate sublines. For example, Satt591 was usedto select sublines from two different elite lines (93B01 and 93M90).Breeding Bias analysis alone indicated that Satt591 allele 3 was the onefavored by breeders over time. In the case of 93B01, allele 3 was thefavorable allele since it was the genotype of the better-yieldingsublime. In contrast, in the ease of 93M90, marker allele 1 was thefavorable allele.

For example, while the Breeding Bias analysis identifies marker locilinked to genetic elements which have been favorable on most geneticbackgrounds in a variety of growing environments, epistatis and othernon-additive interactions influence which allele is “favorable” withinspecific populations, or for particular environments. In addition,disease resistance genes, which contribute to higher relative yield whenthe disease is prevalent, have been documented to result in lower yieldin the absence of disease pressure.

Recombination between a marker locus and the linked genetic elementcontributing to improved yield can also reduce efficiency of markerassisted. An accepted and proven genetic principle is that the frequencyof crossing over between two genetic loci, e.g., a marker locus and aquantitative trait locus, is a function of genetic distance between thetwo loci. The only way to avoid such phase reversals is to develop“perfect” markers that are diagnostic of the DNA polymorphism that isresponsible for the phenotypic difference controlled by the QTL. Thatis, recombination can only be eliminated by cloning the QTL, andidentifying the mutation causally determining the difference inphenotype. Development of perfect markers is possible but is not atrivial exercise. It requires DNA sequencing of the surrounding genomicregion and exhaustive sequence-phenotype association to determineconclusively which DNA polymorphism is always associated with thedesired phenotype. This is an expensive and time-consuming endeavor, thebenefits of which can, in large part, be achieved using the methods andmarker loci of the present invention, without the expense and delay ofcloning each significant yield QTL associated with a marker locusidentified using breeding bias. By periodically confirming marker andphenotype association, using the methods of the present invention,breeders can still reap the benefits of linked (but non-perfect)markers.

Breeding Bias is an effective method for identifying genomic regionsthat have undergone directional selection. If a marker is close enough(typically within about 10 CM) to an important QTL in a given ancestor,the marker allele originally linked in coupling to the favorable QTLallele will remain in coupling phase for a sufficient period ofselection under standard breeding procedures to detect that selection isoccurring in the genomic region including the marker. Thus, the markerloci enumerated herein are associated with, and stand as proxies for,QTL contributing to increased yield. Although, with repeated cycles ofrecombination among members of a given gene pool, genetic crossoversbetween the marker locus and the QTL will tend to accumulate andeventually result in a state of “linkage equilibrium” between the markeralleles and the QTL, periodic reassessment using the near isogenicsubline procedures described herein can insure that selection proceedsfor the allele in linkage phase with the desired QTL allele, despite thepotential for recombination.

The above subline experiments indicate that marker-based subliming is aneffective method for purification and improvement of elite soybeanlines. If the number of markers segregating within a given line orpopulation is small, non-additive effects and linkage phase (coupling orrepulsion) do not pose a problem, as it is fairly inexpensive to fieldtest all possible recombinants and identify those with the optimalphenotype.

Example 5 Allele Confirmation Using Random Crosses

To increase efficiency of marker assisted selection for improved yieldusing, e.g., the marker loci enumerated herein, the allele of the markerlocus segregating with yield can be confirmed. Following identificationof marker loci by Breeding Bias, a limited number of crosses isperformed between the highest yielding elite parents for a particulargeographic zone or growing environment. Preferably, the parents shouldbe as polymorphic as possible at the identified marker loci. Progeny(F1) from these elite by elite crosses is inbred to generate a largepopulation (e.g., between about 200-5000, typically at least about 1000)F3-derived lines. If desired, inbreeding to later generations can alsobe done to increase genetic variation among lines.

A subset of “tester lines” is randomly selected from among the inbredlines derived from each cross. For example, between about 10 and 500lines can be selected. Typically, between about 50 and 100 inbred linesare randomly selected, and enough seed to conduct a reliable yield trialis produced. Several (i.e., between at least 5 and 12, e.g., 8) plantsfrom each inbred line are genotyped at marker loci segregating in theelite parents from which the line was founded, to determine whether theline is segregating or fixed (homozygous) with respect to the relevantmarker. The remaining lines (“remnant population”) can be stored underconditions that preserve seed viability. If desired, additional linescan be selected for testing, or genotyped for presence of the allelesconfirmed to be in coupling linkage phase.

For each cross, a replicated yield trial of each tester line isperformed. The test is replicated in enough environments to adequatelysample the geographic region of interest and to gain a reliable measureof phenotype. The effect on yield for each marker locus within eachcross is determined by comparing the mean yield of lines with a firstallele to the mean yield of lines with the alternate allele. If thedifference in yield is not significant, the marker can be eliminated inthat cross. In contrast, if the difference in yield is significant, the“favorable” allele is confirmed and the locus is used for subsequentmarker assisted selection for yield.

Confirmation of the favorable alleles also permits identification of a“target genotype” including all of the favorable alleles across allsegregating loci in a particular elite by elite cross. As indicatedabove, the entire remnant population can be screened with the subset ofconfirmed markers to identify those segregants that have the highestnumber of favorable alleles. Typically, at least 5% of the remnant linesthat most closely approach the target genotype will be selected,although additional lines can be included at the breeder's discretion.The selected lines can then be evaluated in highly replicated yieldtests to identify which crosses perform better than either elite parentunder a variety of environments and growing conditions.

While the foregoing invention has been described in some detail forpurposes of clarity and understanding, it will be clear to one skilledin the art from a reading of this disclosure that various changes inform and detail can be made without departing from the true scope of theinvention. For example, all the techniques and apparatus described abovecan be used in various combinations. All publications, patents, patentapplications, and/or other documents cited in this application areincorporated by reference in their entirety for all purposes to the sameextent as if each individual publication, patent, patent application,and/or other document were individually indicated to be incorporated byreference for all purposes.

TABLE 1 Elite Soybean Lines by Geographic Region Central Iowa RF SJ NPHL ME (Iowa) (2003) Canada (North) (Illinois) (East) (MidSouth) (South)91B91 92B05 90A07 90B73 92B38 92B52 93B66 94B73 92B05 92B12 90B11 91B1292B63 91B91 93B67 94B74 92B38 92B38 90B31 91B33 92B74 92B05 93B82 94M7092B52 92B52 90B43 91B53 93B01 92B38 93B84 94M90 92B63 92B63 90B72 91B9193B11 92B63 93B85 95B32 92B74 92B74 90B73 91B92 93B15 92B74 93B86 95B3392B75 92M30 91B01 92B05 93B41 93B01 93B87 95B34 92B84 92M31 91B12 92B1293B66 93B46 94B01 95B42 93B01 92M70 91B33 92B23 93B67 93B67 94B23 95B4393B41 92M71 91B52 92B38 93B82 93B82 94B24 95B53 93B45 92M72 91B53 92B6393B85 93B86 94B53 95B95 93B47 92M80 91B64 92B74 93B86 93B87 94B54 95B9693B66 92M91 92B95 A1395 93B87 A3431 94B73 95B97 93B67 93B09 93B25 CX10594B53 CX289 A4009 95M80 93B72 93B36 93B26 D00566D362 94B54 CX394C A413896B21 93B82 93B41 A1395 EX04C00 94B73 EX39E00 A4415 96B32 93B85 93B45CX105 EX06A00 CM428 G3362 A4595 96B51 93B86 93B66 EX10F01 EX10F01MO15733 MO15733 A4715 96M20 93B87 93B67 P1677 EX13P01 MP39009 P9273CM428 97B52 94B23 93B68 P9007 EX13Q01 P9306 P9281 CX469C 97B61 94B5493B82 P9008 EX15N01 P9395 P9306 P9395 A5403 A2835 93B85 P9041 EX16N00S32Z3 P9392 P9481 A5560 A2943 93B86 P9042 EX16P01 S33N1 P9395 P9482A5843 A3127 93B87 P9061 EX22Y01 S38T8 S3911 P9492 A5885 A3242 93M10P9062 EX22Z01 S42H1 ST2660 S3911 A5979 CX232 93M30 P9063 JIM ST2660ST3380 S4260 A5980 CX253 93M40 P9071 KORADA ST2870 ST3883 S42H1 A6297HS93-4118 93M50 P9092 MO400644-02 ST3171 XB31C01 S43B5 BEDFORD P927393M60 P9132 MO413735- ST3630 XB33B ST3870 CLIFFORD 11-52 P9281 93M80P9141 MO501577- ST3883 XB34F01 ST3883 CM428 27-23 P9305 93M90 P9163MO505469- XB31C01 XB35D XB41M01 DAVIS 61-89 P9306 93M92 P9182 P9007XB34F01 XB35W00 XB42J00 ESSEX P9321 93M93 P9203 P9151 XB38A01 XB38A01XB42M01 EX53F03 P9341 A2722 P9244 P9233 XB42M01 YB25Y01 XB48H01 EX56H03P9395 A3237 R01154 S03W4 XB48H01 YB25Z01 YB40M01 EX61E03 R002 S22C3A3322 S0066 S0880 YB28N01 YB27X01 YB40N01 FORREST ST1970 A3431 S0880S19T9 YB29H01 YB27Y01 YB41Q01 FOWLER ST2250 A4138 S1550 S20F8 YB29J01YB28N01 YB48L01 HARTWIG ST2488 EX23B03 S1990 S24L2 YB30J01 YB29H01HOLLADAY ST2688 EX34T03 S19T9 S25J5 YB30N01 YB29J01 HOOD ST2870 EX35F03ST1073 ST0653 YB35C01 YB30J01 HUTCHESON ST3171 EX36Y01 XB03F01 ST1090YB36V00 YB30P01 LEE ST3380 EX40T03 XB07E01 ST1970 YB39M01 YB33K01MANOKIN ST3630 EX44V03 TRAILL YB40M01 YB35C01 P9481 ST3870 S25J5 X9916YB40N01 YB36V00 P9482 ST3883 S32Z3 XB03F01 YB48L01 YB40N01 P9492 XB22R01ST1570 XB10D01 P9521 XB25W01 ST1690 XB15M01 P9552 XB31C01 ST1970 XB20M01P9561 XB34F01 ST2488 XB22R01 P9584 XB35W00 ST2686 XB25W01 P9591 XB38A01ST2788 YB03E00 P9592 XB42M01 ST2870 YB03G01 P9593 YB22W01 ST3171 YB08D01P9594 YB25R99 ST3630 YB09F01 P9611 YB25Y01 ST3660 YB09G01 P9631 YB27X01ST3870 YB10E01 P9641 YB28N01 ST3883 YB11D01 P5960 YB29H01 XB19U04YB14H01 PHARAOH YB29J01 XB22C04 YB15K99 RA451 YB30J01 XB23W03 YB21F01S5960 YB30P01 XB23Y02 YB21G01 S6189 YB31E01 XB25E02 YB22S00 S6262YB32K01 XB25L04 YB22V01 TRACY YB33K01 XB25X04 YB22W01 XB48H01 YB34H01XB26L04 YB22X01 XB48T04 YB37A01 XB27P04 YB24Z01 XB53J04 YB39M01 XB29A04YB25R99 XB54K01 YB40M01 XB29D01 YB25X00 XB55J01 YB40N01 XB29K04 YB25Y01XB55K04 YB41Q01 XB29L04 YB25Z01 XB57M04 XB30E04 YB27Y01 XB58P99 XB31R04XB58Y02 XB34D04 XB58Z04 XB35L04 XB59U04 YB25R03 XB63D00 YB27L03 XB64C04YB27S00 XB67A00 YB28A03 YB48L01 YB29T04 YB51D00 YB34R03 YB52J00 YB34S03YB53C00 YB36E03 YB53D00 YB38E03 YB53E00 YB38G03 YB53G03 YB39V03 YB53H03YB53K03 YB54H00 YB54J00 YB54L00 YB55G00 YB55H00 YB56E00 YB56G03 YB57L03YB58B04 YB59T03 YB59V03 YB60N01 YB60P03 YB63E00 YB65C00 YOUNG

TABLE 2 Exemplary Target Progeny from Segregating Cross. Target MarkerParent 1 Parent 2 Progeny  1 + − +  2 − + +  3 + − +  4 − + +  5 + − + 6 − + +  7 + − +  8 − + +  9 + − + 10 − + + where + = contains the mostfavorable allele − = contains some other allele

TABLE 3 Favorable Allelic Forms: Central Region Region ChromosomePosition Locus Allele Sig. 2 A1 19.1 SATT042 3 ** 2 A1 19.1 SATT364 3 **2 A1 19.1 SATT454 1 ** 2 A1 19.1 SATT526 1 ** 3 A1 27.1 SATT300 3 ** 3A1 27.1 SATT591 3 ** 3 A1 28.5 SATT155 2 ** 4 A1 69.9 SATT385 1 * 5 A187.3 SATT236 1 * 5 A1 87.3 SATT511 1 * 6 A2 0.0 P12390B-1 1 ** 13 A2184.0 SATT429 4 ** 15 B1 39.0 SATT197 5 * 17 B1 74.1 SATT583 1 * 20 B220.0 P12105A-1 1 * 21 B2 45.0 P10641A-1 2 ** 23 B2 86.1 SATT556 2 * 23B2 88.6 SATT020 2 * 25 B2 110.9 SATT534 7 * 26 B2 120.0 P10638B-2 1 * 29C1 95.8 SATT399 3 ** 29 C1 96.5 SATT361 2 ** 29 C1 98.0 P10639A-1 1 **29 C1 99.0 SATT190 4 ** 31 C1 173.0 SATT338 3 * 36 C2 140.9 SATT557 5 **36 C2 145.8 SATT319 1 * 37 C2 165.8 SATT460 5 * 37 C2 170.5 SATT433 3 *38 C2 193.5 SATT357 1 * 40 D1a 54.7 SATT321 2 * 41 D1a 68.6 SATT383 3 *41 D1a 69.8 SATT295 2 * 42 D1a 80.3 SATT507 3 * 43 D1a 129.2 SATT129 5** 43 D1a 129.2 SATT147 1 * 44 D1b 0.0 SATT216 4 * 45 D1b 20.0 P10621B-22 * 46 D1b 39.8 SATT558 3 * 49 D1b 76.1 SATT546 4 * 51 D1b 120.0P13072A-1 2 * 53 D2 66.9 SATT582 2 ** 54 D2 93.8 SATT389 6 ** 55 D2110.4 SATT464 1 ** 57 D2 148.3 SATT413 3 * 60 E 40.0 P13074A-1 1 * 62 E98.0 SATT573 2 * 62 E 98.0 SATT598 1 ** 62 E 100.6 SATT263 2 * 62 E101.0 SATT602 4 * 62 E 102.2 SATT151 4 * 62 E 102.2 SATT355 3 * 62 E102.2 SATT452 2 * 64 F 0.0 SATT146 1 ** 64 F 0.0 SATT193 3 ** 64 F 0.0SATT569 3 ** 64 F 1.7 SATT343 3 ** 64 F 1.9 SATT586 5 ** 66 F 57.5SATT595 1 * 67 F 85.0 P10782A-1 1 ** 67 F 95.0 SATT334 3 * 70 F 157.8SATT144 1 * 71 F 165.8 SATT522 5 * 73 G 8.5 SATT356 2 * 75 G 66.1SATT533 1 * 77 G 100.8 SATT199 1 ** 77 G 103.2 SATT503 3 * 77 G 103.2SATT517 4 * 79 G 137.0 SATT191 2 * 79 G 138.4 SAT_117 1 * 81 H 0.0SATT353 2 * 85 H 125.3 SATT181 5 ** 86 I 15.5 SATT127 2 * 87 I 57.9SATT270 5 * 92 J 61.3 SCT_065 2 ** 92 J 63.6 SATT596 4 ** 92 J 68.0SATT406 2 ** 92 J 70.8 SATT380 3 ** 92 J 72.6 SATT183 1 ** 92 J 74.4SATT529 1 ** 94 K 17.9 SATT242 4 ** 96 K 70.9 SATT441 2 * 96 K 72.8SATT544 2 * 97 K 85.8 SATT240 2 * 101 L 34.6 SATT398 4 ** 102 L 42.3SATT497 3 * 102 L 47.7 SATT284 2 * 103 L 77.1 SATT166 3 ** 103 L 78.0SATT448 4 * 104 L 118.0 SATT373 3 ** 104 L 118.6 SATT513 11 * 104 L124.5 P12394A-1 2 ** 108 M 87.6 SATT536 4 ** 108 M 91.1 SATT175 5 * 109M 115.0 P10615A-1 1 * 110 M 143.5 SATT346 3 * 111 M 173.5 SATT336 4 *112 N 20.0 P13069A-1 3 ** 112 N 25.0 P5467A-1 2 ** 115 N 88.6 SATT3395 * 118 O 2.4 P12396A-1 1 * 118 O 6.3 SATT487 4 ** 119 O 37.7 SATT259 4** 120 O 60.5 SATT420 2 ** 120 O 65.1 SATT576 4 * 122 O 103.8 SATT4773 * 123 O 125.0 SATT581 2 ** 124 O 153.3 SATT153 3 ** 124 O 155.1SATT243 1 ** 128 UM # P10793A-1 2 * 129 UM # P12391A-1 1 * 129 UM #P12392A-1 1 * 131 UM # P13560A-1 1 * 131 UM # P13561A-1 2 ** 133 UM #SAC1677 3 ** 134 UM # SATT040 4 ** 139 UM # SATT111 3 ** 140 UM #SATT176 3 ** 143 UM # SATT299 5 *

TABLE 4 Favorable Allelic Forms: Canada Region Chromosome Position LocusAllele Sig. 4 A1 69.9 SATT385 6 * 5 A1 87.3 SATT225 1 * 9 A2 108.7SATT508 2 ** 11 A2 136.0 P10635A-1 1 * 15 B1 39.0 SATT197 5 * 17 B1 68.1SATT597 3 * 17 B1 71.6 SCT_026 1 * 17 B1 73.3 SATT415 3 * 17 B1 74.1SATT583 1 * 18 B1 80.0 P12198A-1 1 * 18 B1 92.1 SATT359 1 * 23 B2 86.1SATT556 2 * 29 C1 95.8 SATT399 3 ** 29 C1 96.5 SATT361 2 * 29 C1 98.0P10639A-1 1 ** 29 C1 99.0 SATT190 4 ** 31 C1 173.0 SATT338 3 ** 32 C225.4 SATT227 1 * 33 C2 48.1 SATT422 4 * 36 C2 145.8 SATT319 3 * 41 D1a68.6 SATT267 1 ** 42 D1a 80.3 SATT507 3 ** 42 D1a 85.0 P10620A-1 1 * 43D1a 129.2 SATT129 2 * 46 D1b 39.8 SATT558 3 * 48 D1b 65.9 SATT506 3 * 53D2 66.9 SATT582 2 ** 55 D2 110.4 SATT464 1 ** 62 E 100.6 SATT204 4 * 62E 100.6 SATT263 2 * 62 E 101.0 SATT491 3 ** 62 E 102.2 SATT151 4 * 62 E102.2 SATT355 3 * 62 E 102.2 SATT452 2 * 64 F 0.0 SATT146 1 ** 64 F 0.0SATT193 3 * 64 F 0.0 SATT569 3 * 64 F 1.7 SATT343 3 * 64 F 1.7 SATT3435 * 64 F 1.9 SATT586 5 ** 67 F 85.0 P10782A-1 1 ** 67 F 90.0 P10598A-11 * 67 F 95.0 SATT334 3 ** 68 F 114.8 SATT510 3 * 72 F 175.0 P9026A-12 * 75 G 53.0 SATT115 1 * 85 H 125.3 SATT181 5 ** 86 I 9.8 SATT367 2 *86 I 15.5 SATT127 2 ** 86 I 16.6 SCTT012 2 * 87 I 57.9 SATT270 5 * 89 I114.2 SATT440 2 * 92 J 68.0 SATT406 2 * 92 J 70.8 SATT380 3 * 92 J 72.6SATT183 1 ** 92 J 74.4 SATT529 1 * 96 K 70.9 SATT441 3 * 97 K 85.8SATT240 2 * 101 L 32.4 SATT523 1 * 103 L 77.1 SATT166 3 * 103 L 78.0SATT448 1 * 104 L 124.5 P12394A-1 2 ** 108 M 87.6 SATT536 4 * 110 M143.5 SATT346 3 * 112 N 20.0 P13069A-1 2 ** 113 N 35.4 SATT584 3 * 113 N40.0 P3050A-2 2 * 114 N 61.2 SATT387 4 * 118 O 2.4 P12396A-1 1 * 118 O6.3 SATT487 4 ** 119 O 37.7 SATT259 4 * 119 O 39.0 SATT347 2 * 120 O69.2 SATT262 5 * 123 O 130.0 P11070A-1 1 ** 131 UM # P13560A-1 1 ** 131UM # P13561A-1 2 ** 133 UM # SAC1677 3 * 134 UM # SATT040 4 * 138 UM #SATT109 3 ** 139 UM # SATT111 3 ** 140 UM # SATT176 3 * 140 UM # SATT1765 * 145 UM # SATT512 5 **

TABLE 5 Favorable Allelic Forms: North Region Region Chromosome PositionLocus Allele Sig. 2 A1 19.1 SATT454 1 * 2 A1 19.1 SATT526 1 * 3 A1 27.1SATT300 3 * 3 A1 27.1 SATT591 3 * 6 A2 0.0 P12390B-1 1 * 9 A2 108.7SATT327 2 * 9 A2 108.7 SATT508 2 * 12 A2 154.7 SATT409 5 ** 13 A2 184.0SATT429 4 ** 14 B1 22.5 SATT426 5 * 14 B1 26.7 SATT509 1 ** 15 B1 39.0SATT197 5 ** 17 B1 73.3 SATT415 3 * 17 B1 74.1 SATT583 1 * 18 B1 80.0P12198A-1 1 ** 18 B1 85.0 P8584A-1 1 * 18 B1 92.1 SATT359 1 * 21 B2 45.0P10641A-1 2 * 23 B2 86.1 SATT556 2 * 23 B2 86.1 SATT556 4 * 23 B2 88.6SATT020 2 * 25 B2 110.9 SATT534 7 * 29 C1 95.8 SATT399 3 ** 29 C1 96.5SATT361 2 ** 29 C1 98.0 P10639A-1 1 ** 29 C1 99.0 SATT190 4 ** 31 C1173.0 SATT338 3 ** 32 C2 25.4 SATT227 1 ** 33 C2 53.4 SATT457 1 * 36 C2140.9 SATT557 5 * 37 C2 168.3 P13073A-1 2 * 37 C2 170.5 SATT433 3 * 41D1a 68.6 SATT267 1 * 42 D1a 80.3 SATT507 3 ** 42 D1a 82.0 SAT_110 3 * 43D1a 129.2 SATT129 5 * 46 D1b 39.8 SATT558 3 * 48 D1b 65.9 SATT506 3 **51 D1b 120.0 P13072A-1 2 * 53 D2 66.9 SATT582 2 ** 54 D2 93.8 SATT389 6** 54 D2 99.6 SATT461 1 * 55 D2 110.4 SATT464 1 ** 62 E 98.0 SATT573 2 *62 E 98.0 SATT598 1 * 62 E 100.6 SATT204 3 * 64 F 0.0 SATT146 1 ** 64 F0.0 SATT193 3 ** 64 F 0.0 SATT569 3 ** 64 F 1.7 SATT343 3 ** 64 F 1.9SATT586 5 ** 65 F 18.0 SATT348 2 * 67 F 85.0 P10782A-1 1 ** 67 F 90.0P10598A-1 1 * 67 F 95.0 SATT334 3 * 70 F 157.8 SATT144 1 * 71 F 165.8SATT522 4 * 75 G 53.0 SATT115 3 * 75 G 61.3 SATT594 5 * 76 G 71.6SATT303 6 * 76 G 72.4 SATT566 1 * 77 G 100.8 SATT199 1 * 77 G 103.2SATT517 2 * 81 H 0.0 SATT353 2 * 85 H 125.3 SATT181 5 ** 86 I 15.5SATT127 2 ** 87 I 57.9 SATT270 5 ** 89 I 114.2 SATT440 2 ** 90 J 10.5SATT249 4 * 92 J 61.3 SCT_065 2 * 92 J 63.6 SATT596 4 * 92 J 68.0SATT406 2 ** 92 J 70.8 SATT380 3 ** 92 J 72.6 SATT183 1 ** 92 J 74.4SATT529 1 ** 94 K 17.9 SATT242 4 * 101 L 32.4 SATT523 1 * 101 L 34.6SATT398 4 ** 102 L 42.3 SATT497 3 ** 103 L 77.1 SATT166 3 ** 103 L 78.0SATT448 4 * 104 L 118.6 SATT513 11 * 104 L 124.5 P12394A-1 2 ** 108 M87.6 SATT536 4 ** 108 M 91.1 SATT175 5 * 110 M 143.5 SATT346 3 * 111 M173.5 SATT336 4 * 112 N 20.0 P13069A-1 3 ** 113 N 38.3 SAT_084 2 * 115 N88.6 SATT339 5 * 116 N 92.2 SATT255 3 * 118 O 2.4 P12396A-1 1 ** 118 O6.3 SATT487 4 ** 119 O 37.7 SATT259 4 ** 120 O 60.5 SATT420 2 * 120 O65.1 SATT576 4 * 123 O 130.0 P11070A-1 1 * 129 UM # P12391A-1 1 * 131 UM# P13560A-1 1 ** 131 UM # P13561A-1 2 ** 133 UM # SAC1677 3 ** 134 UM #SATT040 4 ** 138 UM # SATT109 3 * 139 UM # SATT111 3 ** 140 UM # SATT1763 **

TABLE 6 Favorable Allelic Forms: Illinois Region Chromosome PositionLocus Allele Sig. 2 A1 19.1 SATT042 3 ** 2 A1 19.1 SATT364 3 ** 2 A119.1 SATT454 1 ** 2 A1 19.1 SATT526 1 ** 3 A1 27.1 SATT300 3 ** 3 A127.1 SATT591 3 ** 3 A1 28.5 SATT155 2 ** 4 A1 69.9 SATT385 1 * 5 A1 87.3SATT511 1 * 6 A2 0.0 P12390B-1 1 ** 13 A2 184.0 SATT429 4 ** 15 B1 39.0SATT197 5 * 16 B1 56.6 SATT519 1 * 21 B2 45.0 P10641A-1 2 ** 23 B2 86.1SATT556 2 * 23 B2 87.0 SATT272 1 * 23 B2 88.6 SATT020 2 * 24 B2 97.3SATT066 3 * 26 B2 120.0 P10638B-2 1 * 29 C1 95.8 SATT399 3 ** 29 C1 96.5SATT361 2 ** 29 C1 98.0 P10639A-1 1 ** 29 C1 99.0 SATT190 4 ** 31 C1173.0 SATT338 3 * 36 C2 140.9 SATT557 5 ** 36 C2 145.8 SATT319 1 * 37 C2165.8 SATT460 5 * 37 C2 170.5 SATT433 3 * 38 C2 193.5 SATT357 1 * 40 D1a54.7 SATT321 2 * 42 D1a 73.1 SATT203 4 * 43 D1a 129.2 SATT129 5 ** 43D1a 129.2 SATT147 1 * 44 D1b 0.0 SATT216 4 * 45 D1b 20.0 P10621B-2 2 *46 D1b 39.8 SATT558 3 * 54 D2 93.8 SATT389 6 * 55 D2 110.4 SATT464 1 **60 E 40.0 P13074A-1 1 * 62 E 98.0 SATT573 2 * 62 E 98.0 SATT598 1 * 62 E100.6 SATT263 2 * 62 E 101.0 SATT602 4 * 62 E 102.2 SATT151 4 * 62 E102.2 SATT355 3 * 62 E 102.2 SATT452 2 * 64 F 0.0 SATT146 1 * 64 F 0.0SATT193 3 ** 64 F 0.0 SATT569 3 ** 64 F 1.7 SATT343 3 ** 64 F 1.9SATT586 5 ** 66 F 57.5 SATT595 1 ** 67 F 85.0 P10782A-1 1 * 67 F 95.0SATT334 3 * 70 F 157.8 SATT144 1 * 71 F 165.8 SATT522 5 * 73 G 4.0P7659A-2 2 * 73 G 7.7 SATT570 3 ** 73 G 8.5 SATT356 2 * 75 G 53.0SATT115 1 ** 75 G 66.1 SATT533 1 * 77 G 100.8 SATT199 1 ** 77 G 103.2SATT503 3 * 77 G 103.2 SATT517 4 * 79 G 137.0 SATT191 2 ** 79 G 138.4SAT_117 1 ** 83 H 77.3 SATT279 6 * 85 H 125.3 SATT181 5 ** 86 I 15.5SATT127 2 * 92 J 61.3 SCT_065 2 ** 92 J 63.6 SATT596 4 * 92 J 68.0SATT406 2 ** 92 J 70.8 SATT380 3 ** 92 J 72.6 SATT183 1 ** 92 J 74.4SATT529 1 ** 94 K 17.9 SATT242 4 ** 97 K 85.8 SATT240 2 * 101 L 34.6SATT398 4 ** 102 L 42.3 SATT497 3 * 103 L 77.1 SATT166 3 ** 103 L 78.0SATT448 4 * 104 L 118.0 SATT373 3 ** 104 L 124.5 P12394A-1 2 ** 105 M12.4 SATT590 17 * 106 M 41.0 SATT567 3 * 107 M 78.9 SATT220 3 * 108 M87.6 SATT536 4 * 108 M 91.1 SATT175 5 * 109 M 115.0 P10615A-1 1 * 112 N20.0 P13069A-1 3 ** 112 N 25.0 P5467A-1 2 ** 112 N 25.0 P5467A-2 1 * 113N 38.3 SAT_084 2 * 115 N 88.6 SATT339 5 * 118 O 2.4 P12396A-1 1 * 118 O6.3 SATT487 4 ** 119 O 37.7 SATT259 4 * 120 O 60.5 SATT420 2 ** 120 O65.1 SATT576 4 * 122 O 103.8 SATT477 3 * 123 O 125.0 SATT581 2 ** 123 O130.0 P11070A-1 1 * 124 O 153.3 SATT153 3 ** 124 O 155.1 SATT243 1 * 127UM # P10632A-1 2 * 128 UM # P10793A-1 2 * 129 UM # P12391A-1 1 * 129 UM# P12392A-1 1 * 131 UM # P13560A-1 1 * 131 UM # P13561A-1 2 ** 133 UM #SAC1677 3 ** 134 UM # SATT040 4 ** 139 UM # SATT111 3 ** 140 UM #SATT176 3 ** 141 UM # SATT219 1 * 143 UM # SATT299 5 *

TABLE 7 Favorable Allelic Forms: East Region Region Chromosome PositionLocus Allele Sig. 2 A1 19.1 SATT042 3 ** 2 A1 19.1 SATT364 3 ** 2 A119.1 SATT454 1 ** 2 A1 19.1 SATT526 1 ** 3 A1 27.1 SATT300 3 ** 3 A127.1 SATT591 3 ** 3 A1 28.5 SATT155 2 ** 4 A1 69.9 SATT385 1 * 5 A1 87.3SATT236 1 * 5 A1 87.3 SATT511 1 * 6 A2 0.0 P12390B-1 1 ** 12 A2 154.7SATT409 5 * 13 A2 184.0 SATT429 4 ** 15 B1 39.0 SATT197 5 * 17 B1 74.1SATT583 1 * 21 B2 45.0 P10641A-1 2 ** 23 B2 86.1 SATT556 2 * 23 B2 88.6SATT020 2 * 26 B2 120.0 P10638B-2 1 * 29 C1 95.8 SATT399 3 ** 29 C1 96.5SATT361 2 ** 29 C1 98.0 P10639A-1 1 ** 29 C1 99.0 SATT190 4 ** 31 C1173.0 SATT338 3 * 36 C2 140.9 SATT557 5 ** 36 C2 145.8 SATT319 1 * 37 C2170.5 SATT433 3 * 38 C2 193.5 SATT357 1 * 40 D1a 54.7 SATT321 2 * 41 D1a68.6 SATT383 3 * 43 D1a 129.2 SATT129 5 ** 43 D1a 129.2 SATT147 1 * 45D1b 20.0 P10621B-2 2 * 46 D1b 39.8 SATT558 3 * 53 D2 66.9 SATT582 2 * 54D2 93.8 SATT389 6 * 55 D2 110.4 SATT464 1 ** 60 E 40.0 P13074A-1 1 * 62E 98.0 SATT573 2 * 62 E 98.0 SATT598 1 * 62 E 100.6 SATT263 2 * 62 E101.0 SATT602 4 * 62 E 102.2 SATT151 4 * 62 E 102.2 SATT355 1 * 62 E102.2 SATT452 2 * 64 F 0.0 SATT146 1 ** 64 F 0.0 SATT193 3 ** 64 F 0.0SATT569 3 ** 64 F 1.7 SATT343 3 ** 64 F 1.9 SATT586 5 ** 66 F 57.5SATT595 1 * 67 F 85.0 P10782A-1 1 ** 67 F 95.0 SATT334 3 * 70 F 157.8SATT144 1 * 73 G 8.5 SATT356 2 * 75 G 53.0 SATT115 1 ** 75 G 66.1SATT533 1 * 77 G 100.8 SATT199 1 ** 77 G 103.2 SATT503 3 * 77 G 103.2SATT517 4 ** 79 G 137.0 SATT191 2 * 79 G 138.4 SAT_117 1 * 83 H 77.3SATT279 6 * 85 H 125.3 SATT181 5 ** 86 I 15.5 SATT127 2 * 92 J 61.3SCT_065 2 ** 92 J 63.6 SATT596 4 ** 92 J 68.0 SATT406 2 ** 92 J 70.8SATT380 3 ** 92 J 72.6 SATT183 1 ** 92 J 74.4 SATT529 1 ** 94 K 17.9SATT242 4 ** 97 K 85.8 SATT240 2 * 103 L 77.1 SATT166 3 ** 104 L 118.0SATT373 3 ** 104 L 124.5 P12394A-1 2 ** 108 M 87.6 SATT536 4 ** 108 M91.1 SATT175 5 * 109 M 115.0 P10615A-1 1 * 110 M 143.5 SATT346 3 * 111 M173.5 SATT336 4 * 112 N 20.0 P13069A-1 3 ** 112 N 25.0 P5467A-1 2 ** 112N 25.0 P5467A-2 1 * 113 N 38.3 SAT_084 2 * 115 N 88.6 SATT339 5 * 118 O2.4 P12396A-1 1 * 118 O 6.3 SATT487 4 ** 119 O 37.7 SATT259 4 ** 120 O60.5 SATT420 2 ** 120 O 65.1 SATT576 4 * 120 O 69.2 SATT473 2 * 122 O103.8 SATT477 3 * 123 O 125.0 SATT581 2 ** 123 O 130.0 P11070A-1 1 * 124O 153.3 SATT153 3 ** 124 O 155.1 SATT243 1 ** 128 UM # P10793A-1 2 * 129UM # P12391A-1 1 * 129 UM # P12392A-1 1 * 131 UM # P13560A-1 1 * 131 UM# P13561A-1 2 ** 133 UM # SAC1677 3 ** 134 UM # SATT040 4 ** 139 UM #SATT111 3 ** 140 UM # SATT176 3 ** 141 UM # SATT219 9 ** 143 UM #SATT299 5 *

TABLE 8 Favorable Allelic Forms: Mid South Region Region ChromosomePosition Locus Allele Sig. 2 A1 19.1 SATT042 3 * 2 A1 19.1 SATT364 3 **2 A1 19.1 SATT454 1 ** 2 A1 19.1 SATT526 1 ** 3 A1 27.1 SATT300 3 ** 3A1 27.1 SATT591 3 ** 3 A1 28.5 SATT155 2 ** 5 A1 87.3 SATT511 1 * 6 A20.0 P12390B-1 1 * 8 A2 96.2 SATT233 6 * 11 A2 136.0 P10635A-1 1 * 12 A2154.7 SATT409 2 * 12 A2 161.8 SATT228 4 ** 13 A2 184.0 SATT429 4 ** 15B1 39.0 SATT197 5 * 18 B1 92.1 SATT359 3 ** 20 B2 20.0 P12105A-1 1 * 21B2 45.0 P10641A-1 2 ** 23 B2 86.1 SATT556 2 * 23 B2 87.0 SATT272 1 * 23B2 88.6 SATT020 2 * 29 C1 95.8 SATT399 3 ** 29 C1 96.5 SATT361 2 * 29 C198.0 P10639A-1 1 * 29 C1 99.0 SATT190 4 ** 31 C1 173.0 SATT338 3 * 36 C2140.9 SATT557 5 ** 36 C2 145.8 SATT319 1 * 37 C2 165.8 SATT460 5 * 40D1a 54.7 SATT321 2 ** 42 D1a 73.1 SATT203 4 * 43 D1a 129.2 SATT129 5 **44 D1b 0.0 SATT216 4 * 45 D1b 20.0 P10621B-2 2 ** 46 D1b 39.8 SATT5583 * 54 D2 93.8 SATT389 6 * 55 D2 110.4 SATT464 1 ** 62 E 98.0 SATT5981 * 62 E 100.6 SATT263 2 * 62 E 102.2 SATT151 4 * 62 E 102.2 SATT355 3 *62 E 102.2 SATT452 2 * 64 F 0.0 SATT146 1 ** 64 F 0.0 SATT193 3 ** 64 F0.0 SATT569 3 ** 64 F 1.7 SATT343 3 ** 64 F 1.9 SATT586 5 ** 65 F 16.4SATT423 3 * 65 F 18.0 SATT348 4 * 66 F 57.5 SATT595 1 * 67 F 85.0P10782A-1 1 ** 67 F 95.0 SATT334 3 * 73 G 2.0 P10646A-1 2 ** 73 G 3.0P5219A-1 2 * 73 G 4.0 P7659A-2 2 * 73 G 7.7 SATT570 3 ** 73 G 8.5SATT356 2 * 75 G 53.0 SATT115 1 * 75 G 66.1 SATT533 1 * 77 G 100.8SATT199 1 * 77 G 103.2 SATT503 3 * 77 G 103.2 SATT517 4 * 79 G 137.0SATT191 2 * 79 G 138.4 SAT_117 1 ** 84 H 116.9 SATT142 3 * 85 H 125.3SATT181 5 ** 86 I 9.8 SATT367 6 * 86 I 15.5 SATT127 2 * 89 I 114.2SATT440 3 * 92 J 61.3 SCT_065 2 ** 92 J 63.6 SATT596 4 * 92 J 68.0SATT406 2 ** 92 J 70.8 SATT380 3 ** 92 J 72.6 SATT183 1 ** 92 J 74.4SATT529 1 ** 94 K 17.9 SATT242 4 * 96 K 72.8 SATT544 2 * 97 K 85.8SATT240 2 * 101 L 34.6 SATT398 4 ** 102 L 42.3 SATT497 3 * 103 L 77.1SATT166 3 ** 103 L 78.0 SATT448 4 * 104 L 124.5 P12394A-1 2 * 107 M 78.9SATT220 3 * 108 M 87.6 SATT536 4 * 109 M 115.0 P10615A-1 1 * 110 M 143.5SATT346 3 * 115 N 88.6 SATT339 5 * 118 O 2.4 P12396A-1 1 * 118 O 6.3SATT487 4 ** 119 O 37.7 SATT259 4 * 120 O 60.5 SATT420 2 * 120 O 65.1SATT576 4 * 122 O 103.8 SATT477 3 * 123 O 125.0 SATT581 2 ** 124 O 153.3SATT153 3 ** 124 O 155.1 SATT243 1 * 129 UM # P12392A-1 1 * 131 UM #P13560A-1 1 * 131 UM # P13561A-1 2 ** 133 UM # SAC1677 3 * 134 UM #SATT040 4 ** 139 UM # SATT111 3 ** 140 UM # SATT176 3 ** 141 UM #SATT219 9 ** 143 UM # SATT299 5 *

TABLE 9 Favorable Allelic Forms: South Region Region Chromosome PositionLocus Allele Sig. 2 A1 14.6 SATT165 2 * 2 A1 19.1 SATT042 3 * 2 A1 19.1SATT042 4 * 2 A1 19.1 SATT364 4 * 2 A1 19.1 SATT526 1 ** 3 A1 27.1SATT300 3 ** 3 A1 27.1 SATT591 3 * 3 A1 28.5 SATT155 2 ** 4 A1 69.9SATT385 1 * 5 A1 87.3 SATT236 1 * 5 A1 87.3 SATT511 1 * 6 A2 0.0P12390B-1 1 * 7 A2 20.0 SATT480 1 * 9 A2 108.7 SATT329 2 * 14 B1 26.7SATT509 6 * 17 B1 71.6 SCT_026 2 ** 17 B1 74.1 SATT583 3 * 17 B1 74.8SATT430 1 * 18 B1 92.1 SATT359 3 ** 19 B1 120.0 P10648A-1 2 * 21 B2 55.8SATT168 2 * 23 B2 86.1 SATT556 2 * 23 B2 87.0 SATT272 2 * 29 C1 95.8SATT399 3 * 29 C1 99.0 SATT190 4 * 33 C2 53.4 SATT457 2 * 37 C2 168.3SATT307 2 * 37 C2 168.3 SCT_028 2 * 37 C2 170.5 SATT433 4 * 38 C2 193.5SATT357 1 * 40 D1a 54.7 SATT321 2 * 46 D1b 39.8 SATT558 3 * 47 D1b 51.6SATT266 1 * 48 D1b 64.3 SATT282 4 * 48 D1b 65.1 SATT537 8 * 54 D2 93.8SATT389 6 * 54 D2 99.6 SATT461 1 * 55 D2 106.6 SATT311 2 * 55 D2 107.8SATT514 1 * 55 D2 110.4 SATT464 1 ** 55 D2 112.1 SATT543 4 ** 56 D2139.1 SATT186 1 * 60 E 40.0 P13074A-1 1 * 61 E 80.0 P10624A-1 2 * 62 E100.6 SATT204 1 * 62 E 101.0 SATT491 3 * 62 E 102.2 SATT151 3 * 62 E102.2 SATT355 3 * 62 E 102.2 SATT452 2 ** 64 F 0.0 SATT146 4 ** 64 F 0.0SATT569 3 * 64 F 1.7 SATT343 5 * 64 F 1.9 SATT586 2 * 67 F 87.5 P3436A-12 ** 67 F 95.0 SATT334 3 * 68 F 114.8 SATT510 5 ** 70 F 157.8 SATT1441 * 73 G 2.0 P10646A-1 2 ** 73 G 3.0 P5219A-1 2 * 73 G 4.0 P7659A-2 2 *73 G 7.7 SATT570 3 ** 73 G 8.5 SATT356 2 * 74 G 13.5 SATT130 2 * 75 G61.3 SATT594 6 ** 75 G 66.1 SATT533 1 * 76 G 71.6 SATT303 7 * 76 G 71.6SATT303 9 ** 76 G 72.4 SATT352 10 ** 76 G 72.4 SATT566 3 * 77 G 100.8SATT199 1 ** 77 G 103.2 SATT503 3 ** 77 G 103.2 SATT517 4 * 79 G 137.0SATT191 2 * 79 G 138.4 SAT_117 1 ** 81 H 0.0 SATT353 2 * 82 H 42.3SATT442 4 ** 83 H 77.3 SATT279 5 ** 83 H 77.3 SATT314 1 ** 86 I 9.8SATT367 6 ** 86 I 15.5 SATT127 2 * 87 I 57.9 SATT270 5 * 89 I 115.0P10640A-1 1 * 92 J 67.2 SATT280 5 ** 92 J 70.8 SATT380 3 ** 92 J 72.6SATT183 2 ** 93 J 118.0 SATT431 3 ** 94 K 17.9 SATT242 1 ** 95 K 44.0SATT102 2 * 98 K 144.0 P10618A-1 1 * 98 K 144.3 SATT475 1 * 99 K 164.8SATT196 4 * 101 L 32.4 SATT523 3 * 101 L 33.9 SATT418 4 * 103 L 77.1SATT166 1 ** 103 L 78.0 SATT448 4 * 104 L 118.0 SATT373 17 * 104 L 118.6SATT513 1 ** 104 L 124.5 P12394A-1 2 * 105 M 12.4 SATT590 2 * 108 M 87.6SATT536 4 * 110 M 139.4 SATT250 1 * 113 N 35.4 SATT584 2 * 114 N 61.2SATT387 5 ** 115 N 80.2 SATT549 5 * 117 N 113.0 SATT257 2 * 118 O 2.4P12396A-1 1 * 118 O 6.3 SATT487 1 ** 119 O 37.7 SATT259 3 ** 119 O 39.0SATT347 2 * 120 O 65.1 SATT576 4 * 120 O 67.5 SATT550 1 * 124 O 155.1SATT243 2 * 125 O 175.7 P8230A-1 2 ** 126 UM # P10623A-1 2 * 129 UM #P12391A-1 2 * 131 UM # P13561A-1 1 * 132 UM # p2481A-1 1 * 133 UM #SAC1677 3 ** 136 UM # SATT108 1 * 139 UM # SATT111 3 * 140 UM # SATT1768 ** 141 UM # SATT219 4 ** 143 UM # SATT299 5 * 145 UM # SATT512 3 *

TABLE 10 Favorable Alleles: Iowa (A) Best Marker in Genomic Genomic LODCHG Region Chromosome Position Locus Allele Region Sig. Iowa EXP IowaOBS Iowa Iowa #ELI Iowa 2 A1 19.1 Satt042 3 ** 2.68 0.13 0.75 0.62 67 2A1 19.1 Satt364 3 ** 3.22 0.05 0.72 0.67 68 2 A1 19.1 Satt454 1 ** 3.700.05 0.74 0.69 69 2 A1 19.1 Satt526 1 Best ** 4.00 0.06 0.79 0.72 68 3A1 27.1 Satt300 3 ** 3.22 0.08 0.77 0.69 64 3 A1 27.1 Satt591 3 Best **3.22 0.13 0.82 0.70 67 3 A1 28.5 Satt155 2 ** 2.96 0.18 0.82 0.64 66 5A1 87.3 Satt511 1 Best * 1.40 0.14 0.57 0.44 61 6 A2 0.0 P12390B-1 1Best ** 2.59 0.32 0.89 0.57 66 13 A2 184.0 Satt429 4 Best ** 4.00 0.020.63 0.61 53 15 B1 39.0 Satt197 5 Best * 1.66 0.22 0.64 0.41 48 21 B245.0 P10641A-1 2 Best ** 3.22 0.34 0.94 0.60 70 25 B2 110.9 Satt534 7Best * 1.47 0.49 0.89 0.40 61 26 B2 120.0 P10638B-2 1 Best * 1.91 0.620.94 0.33 70 29 C1 95.8 Satt399 3 ** 4.00 0.08 0.89 0.81 61 29 C1 96.5Satt361 2 ** 3.05 0.15 0.80 0.65 68 29 C1 98.0 P10639A-1 1 ** 3.22 0.190.86 0.66 63 29 C1 99.0 Satt190 4 Best ** 4.00 0.05 0.79 0.73 65 31 C1173.0 Satt338 3 Best * 1.55 0.25 0.71 0.46 63 36 C2 140.9 Satt557 5 Best** 3.52 0.58 1.00 0.42 70 36 C2 145.8 Satt319 1 * 1.90 0.74 1.00 0.26 6737 C2 165.8 Satt460 5 * 1.43 0.63 0.97 0.34 44 37 C2 170.5 Satt433 3Best * 1.59 0.15 0.48 0.34 61 38 C2 193.5 Satt357 1 Best * 1.52 0.680.97 0.29 66 40 D1a 54.7 Satt321 2 Best * 1.82 0.19 0.69 0.50 66 41 D1a69.8 Satt295 2 Best * 1.67 0.70 0.98 0.28 50 42 D1a 80.3 Satt507 3Best * 1.37 0.20 0.63 0.43 66 43 D1a 129.2 Satt129 5 Best ** 2.89 0.310.93 0.61 61 43 D1a 129.2 Satt147 1 * 1.46 0.69 0.97 0.28 62 46 D1b 39.8Satt558 3 Best * 1.76 0.29 0.81 0.52 56 53 D2 66.9 Satt582 2 ** 2.890.01 0.28 0.27 50 54 D2 93.8 Satt389 6 Best ** 2.46 0.05 0.53 0.49 63 55D2 110.4 Satt464 1 Best ** 3.40 0.05 0.67 0.62 66 62 E 98.0 Satt573 2 *1.72 0.46 0.91 0.45 62 62 E 98.0 Satt598 1 Best ** 2.28 0.38 0.92 0.5466 62 E 100.6 Satt263 2 * 1.88 0.07 0.53 0.46 59 62 E 101.0 Satt602 4 *1.36 0.12 0.43 0.31 61 62 E 102.2 Satt151 4 * 1.78 0.05 0.44 0.40 60 64F 0.0 Satt146 1 ** 2.89 0.20 0.86 0.66 50 64 F 0.0 Satt193 3 ** 3.700.10 0.84 0.74 62 64 F 0.0 Satt569 3 ** 3.16 0.15 0.86 0.71 69 64 F 1.7Satt176 3 ** 4.00 0.09 0.85 0.76 71 64 F 1.7 Satt343 3 Best ** 4.00 0.090.89 0.80 66 64 F 1.9 Satt586 5 ** 4.00 0.09 0.84 0.75 55 64 F 2.5Satt040 4 ** 3.52 0.09 0.81 0.73 67 67 F 85.0 P10782A-1 1 Best ** 3.160.27 0.90 0.64 70 70 F 157.8 Satt144 1 Best * 1.79 0.45 0.90 0.45 67 71F 165.8 Satt522 5 Best * 1.48 0.04 0.32 0.27 63 73 G 8.5 Satt356 2Best * 1.42 0.69 0.97 0.28 68 75 G 66.1 Satt533 1 Best * 1.94 0.43 0.900.46 68 77 G 100.8 Satt199 1 Best ** 3.22 0.44 0.98 0.54 65 77 G 103.2Satt517 4 * 1.54 0.10 0.54 0.44 64 79 G 137.0 Satt191 2 * 1.65 0.15 0.620.47 65 79 G 138.4 Sat_117 1 Best * 1.70 0.20 0.68 0.48 66 85 H 125.3Satt181 5 Best ** 2.14 0.05 0.61 0.56 61 86 I 15.5 Satt127 2 Best * 1.990.62 0.99 0.37 58 92 J 61.3 Sct_065 2 ** 3.70 0.26 0.91 0.65 58 92 J63.6 Satt596 4 ** 2.47 0.14 0.79 0.65 65 92 J 68.0 Satt406 2 Best **3.52 0.18 0.87 0.69 66 92 J 70.8 Satt380 3 ** 3.70 0.28 0.94 0.66 63 92J 72.6 Satt183 1 ** 3.30 0.45 0.97 0.53 69 92 J 74.4 Satt529 1 ** 3.520.38 0.97 0.59 61 94 K 17.9 Satt242 4 Best ** 3.05 0.07 0.71 0.64 65 97K 85.8 Satt240 2 Best * 1.49 0.11 0.52 0.41 54 101 L 34.6 Satt398 4 Best** 2.00 0.33 0.83 0.50 51 103 L 77.1 Satt166 3 Best ** 4.00 0.10 0.900.80 61 103 L 78.0 Satt448 4 * 1.46 0.22 0.68 0.46 67 104 L 118.0Satt373 3 ** 3.70 0.00 0.37 0.36 52 104 L 118.6 Satt513 11 * 1.47 0.070.33 0.26 40 104 L 124.5 P12394A-1 2 Best ** 3.05 0.66 1.00 0.35 71 108M 87.6 Satt536 4 Best ** 3.70 0.14 0.83 0.70 66 108 M 91.1 Satt175 5 *1.99 0.19 0.77 0.58 68 109 M 115.0 P10615A-1 1 Best * 1.75 0.10 0.540.44 70 110 M 143.5 Satt346 3 Best * 1.30 0.16 0.61 0.45 61 111 M 173.5Satt336 4 Best * 1.63 0.25 0.71 0.46 44 112 N 20.0 P13069A-1 3 Best **4.00 0.00 0.31 0.31 61 112 N 25.0 P5467A-1 2 ** 4.00 0.00 0.26 0.26 62115 N 88.6 Satt339 5 Best * 1.96 0.19 0.73 0.54 60 118 O 2.4 P12396A-11 * 1.89 0.64 0.97 0.33 66 118 O 6.3 Satt487 4 Best ** 4.00 0.37 0.970.60 64 119 O 37.7 Satt259 4 Best ** 3.30 0.37 0.92 0.55 63 120 O 60.5Satt420 2 Best ** 2.28 0.30 0.85 0.54 68 120 O 65.1 Satt576 4 * 1.800.22 0.77 0.55 44 122 O 103.8 Satt477 3 Best * 1.32 0.43 0.83 0.40 53123 O 125.0 Satt581 2 Best ** 2.70 0.35 0.88 0.53 69 124 O 153.3 Satt1533 Best ** 2.96 0.49 0.94 0.46 68 124 O 155.1 Satt243 1 ** 2.17 0.60 0.960.36 56 129 UM P12391A-1 1 Best * 1.84 0.39 0.84 0.45 69 129 UMP12392A-1 1 Best * 1.30 0.55 0.88 0.33 69 131 UM P13560A-1 1 Best * 1.710.23 0.64 0.41 70 131 UM P13561A-1 2 Best ** 3.10 0.63 1.00 0.37 67 133UM Sac1677 3 Best ** 4.00 0.15 0.90 0.75 70 139 UM Satt111 3 Best **4.00 0.08 0.81 0.74 67 143 UM Satt299 5 Best * 1.59 0.17 0.65 0.48 61 *95% significance level ** 99% significance level

TABLE 11 Significant Alleles at the “Best” marker locus in each genomicregion. SIGNIF LOD EXP OBS CHG #ELI Region Chromosome Position LocusAllele Iowa Iowa Iowa Iowa Iowa Iowa 2 A1 19.1 Satt526 1 ** 4.00 0.060.79 0.72 68 3 A1 27.1 Satt591 3 ** 3.22 0.13 0.82 0.70 67 5 A1 87.3Satt511 1 * 1.40 0.14 0.57 0.44 61 6 A2 0.0 P12390B-1 1 ** 2.59 0.320.89 0.57 66 13 A2 184.0 Satt429 4 ** 4.00 0.02 0.63 0.61 53 15 B1 39.0Satt197 5 * 1.66 0.22 0.64 0.41 48 21 B2 45.0 P10641A-1 2 ** 3.22 0.340.94 0.60 70 25 B2 110.9 Satt534 7 * 1.47 0.49 0.89 0.40 61 26 B2 120.0P10638B-2 1 * 1.91 0.62 0.94 0.33 70 29 C1 99.0 Satt190 4 ** 4.00 0.050.79 0.73 65 31 C1 173.0 Satt338 3 * 1.55 0.25 0.71 0.46 63 36 C2 140.9Satt557 5 ** 3.52 0.58 1.00 0.42 70 37 C2 170.5 Satt433 3 * 1.59 0.150.48 0.34 61 38 C2 193.5 Satt357 1 * 1.52 0.68 0.97 0.29 66 40 D1a 54.7Satt321 2 * 1.82 0.19 0.69 0.50 66 41 D1a 69.8 Satt295 2 * 1.67 0.700.98 0.28 50 42 D1a 80.3 Satt507 3 * 1.37 0.20 0.63 0.43 66 43 D1a 129.2Satt129 5 ** 2.89 0.31 0.93 0.61 61 46 D1b 39.8 Satt558 3 * 1.76 0.290.81 0.52 56 54 D2 93.8 Satt389 6 ** 2.46 0.05 0.53 0.49 63 55 D2 110.4Satt464 1 ** 3.40 0.05 0.67 0.62 66 62 E 98.0 Satt598 1 ** 2.28 0.380.92 0.54 66 64 F 1.7 Satt343 3 ** 4.00 0.09 0.89 0.80 66 67 F 85.0P10782A-1 1 ** 3.16 0.27 0.90 0.64 70 70 F 157.8 Satt144 1 * 1.79 0.450.90 0.45 67 71 F 165.8 Satt522 5 * 1.48 0.04 0.32 0.27 63 73 G 8.5Satt356 2 * 1.42 0.69 0.97 0.28 68 75 G 66.1 Satt533 1 * 1.94 0.43 0.900.46 68 77 G 100.8 Satt199 1 ** 3.22 0.44 0.98 0.54 65 79 G 138.4Sat_117 1 * 1.70 0.20 0.68 0.48 66 85 H 125.3 Satt181 5 ** 2.14 0.050.61 0.56 61 86 I 15.5 Satt127 2 * 1.99 0.62 0.99 0.37 58 92 J 68.0Satt406 2 ** 3.52 0.18 0.87 0.69 66 94 K 17.9 Satt242 4 ** 3.05 0.070.71 0.64 65 97 K 85.8 Satt240 2 * 1.49 0.11 0.52 0.41 54 101 L 34.6Satt398 4 ** 2.00 0.33 0.83 0.50 51 103 L 77.1 Satt166 3 ** 4.00 0.100.90 0.80 61 104 L 124.5 P12394A-1 2 ** 3.05 0.66 1.00 0.35 71 108 M87.6 Satt536 4 ** 3.70 0.14 0.83 0.70 66 109 M 115.0 P10615A-1 1 * 1.750.10 0.54 0.44 70 110 M 143.5 Satt346 3 * 1.30 0.16 0.61 0.45 61 111 M173.5 Satt336 4 * 1.63 0.25 0.71 0.46 44 112 N 20.0 P13069A-1 3 ** 4.000.00 0.31 0.31 61 115 N 88.6 Satt339 5 * 1.96 0.19 0.73 0.54 60 118 O6.3 Satt487 4 ** 4.00 0.37 0.97 0.60 64 119 O 37.7 Satt259 4 ** 3.300.37 0.92 0.55 63 120 O 60.5 Satt420 2 ** 2.28 0.30 0.85 0.54 68 122 O103.8 Satt477 3 * 1.32 0.43 0.83 0.40 53 123 O 125.0 Satt581 2 ** 2.700.35 0.88 0.53 69 124 O 153.3 Satt153 3 ** 2.96 0.49 0.94 0.46 68 129 UMP12391A-1 1 * 1.84 0.39 0.84 0.45 69 129 UM P12392A-1 1 * 1.30 0.55 0.880.33 69 131 UM P13560A-1 1 * 1.71 0.23 0.64 0.41 70 131 UM P13561A-1 2** 3.10 0.63 1.00 0.37 67 133 UM Satt677 3 ** 4.00 0.15 0.90 0.75 70 139UM Satt111 3 ** 4.00 0.08 0.81 0.74 67 143 UM Satt299 5 * 1.59 0.17 0.650.48 61 * 95% significance level ** 99% significance level

TABLE 12 Favorable Alleles: Iowa (B) New Best Marker Marker in inGenomic Genomic Genomic LOD CHG Region Chromosome Position Locus AlleleRegion Region Sig. Iowa EXP Iowa OBS Iowa Iowa #ELI Iowa 1 A1 1.4Satt684 3 NEW Best * 1.84 0.03 0.36 0.36 79 2 A1 19.1 Satt042 3 ** 2.660.14 0.77 0.64 82 2 A1 19.1 Satt364 3 ** 3.16 0.05 0.70 0.65 65 2 A119.1 Satt454 1 ** 3.40 0.06 0.71 0.64 66 2 A1 19.1 Satt526 1 Best **4.00 0.07 0.77 0.70 86 3 A1 27.1 Satt300 3 ** 3.30 0.11 0.79 0.68 82 3A1 27.1 Satt591 3 Best ** 3.40 0.15 0.83 0.69 81 3 A1 28.5 Satt155 2 **2.89 0.18 0.83 0.65 81 4 A1 69.9 Satt385 1 Best * 1.99 0.44 0.91 0.48 807.5 A2 41.8 Satt632-TB 4 NEW Best * 1.88 0.66 1.00 0.34 41 13 A2 184.0Satt429 4 Best ** 4.00 0.04 0.80 0.76 76 14 B1 27.6 SAT_261 1 NEW Best** 2.42 0.49 1.00 0.51 28 21 B2 45.0 P10641A-1 2 Best ** 3.70 0.34 0.990.65 86 23 B2 86.1 Satt556 2 Best * 1.73 0.16 0.75 0.59 76 26 B2 120.0P10638B-2 1 Best * 1.30 0.63 0.93 0.30 85 29 C1 95.8 Satt399 3 ** 4.000.07 0.89 0.82 77 29 C1 96.5 Satt361 2 ** 3.30 0.16 0.83 0.67 64 29 C199.0 SATT661-TB 2 NEW * 1.68 0.39 0.94 0.55 59 29 C1 99.0 Satt190 4 Best** 4.00 0.05 0.83 0.78 79 30 C1 117.6 SAT_311-DB 3 NEW Best * 1.72 0.450.92 0.47 56 31 C1 173.0 Satt338 3 Best * 1.49 0.24 0.67 0.43 47 32 C228.6 SATT640-TB 3 NEW Best * 1.50 0.57 0.98 0.41 45 36 C2 140.9 Satt5575 Best ** 2.68 0.61 1.00 0.28 84 36 C2 145.8 Satt319 1 * 1.66 0.72 1.000.28 84 36.5 C2 153.5 SAT_142-DB 3 NEW Best ** 2.44 0.58 1.00 0.42 62 40D1a 54.7 Satt321 2 Best * 1.52 0.17 0.64 0.47 82 42 D1a 73.1 Satt203 4Best * 1.54 0.07 0.42 0.35 76 43 D1a 129.2 Satt129 5 Best ** 3.52 0.360.99 0.63 75 43 D1a 129.2 Satt147 1 * 1.85 0.70 1.00 0.30 63 45 D1b 19.1SAT_351 3 NEW Best ** 2.08 0.14 0.73 0.59 66 45 D1b 20.0 P10621B-2 2NEW * 1.51 0.60 0.96 0.36 86 46 D1b 36.7 Satt701 2 NEW Best * 1.97 0.150.59 0.44 27 46 D1b 39.8 Satt634 3 NEW * 1.40 0.44 0.89 0.45 27 53 D266.9 Satt582 2 Best * 1.33 0.06 0.37 0.31 78 54 D2 93.8 Satt389 6 Best *1.35 0.11 0.48 0.37 81 55 D2 110.4 Satt464 1 Best ** 4.00 0.05 0.75 0.7083 55 D2 111.8 Satt662 1 NEW ** 2.08 0.06 0.65 0.59 27 57 D2 149.1Satt672 1 NEW Best * 1.64 0.15 0.57 0.42 27 62 E 98.0 Satt573 2 * 1.830.45 0.92 0.47 61 62 E 98.0 Satt598 1 Best * 1.92 0.49 0.95 0.46 73 62 E100.6 Satt263 2 * 1.42 0.11 0.49 0.38 58 62 E 102.2 Satt151 4 * 1.390.08 0.44 0.36 79 62 E 103.7 SAT_273-DB 1 NEW * 1.59 0.11 0.59 0.48 6264 F 0.0 Satt146 1 ** 3.10 0.20 0.89 0.70 75 64 F 0.0 Satt193 3 ** 3.700.09 0.86 0.77 62 64 F 0.0 Satt569 3 ** 3.30 0.18 0.91 0.73 64 64 F 1.7Satt343 3 Best ** 3.70 0.08 0.89 0.81 74 64 F 1.9 Satt586 5 ** 3.10 0.080.82 0.74 70 64 F 2.5 Satt040 4 ** 3.16 0.08 0.81 0.73 62 66 F 57.5Satt595 1 NEW Best ** 2.35 0.69 1.00 0.31 75 67 F 95.0 Satt334 3 NEWBest * 1.55 0.15 0.63 0.48 73 70 F 157.8 Satt144 1 Best * 1.77 0.46 0.930.47 84 71 F 165.8 Satt522 5 Best * 1.32 0.06 0.37 0.31 77 73 G 7.7Satt570 3 NEW Best ** 3.70 0.00 0.27 0.27 82 73 G 8.5 Satt356 2 Best *1.30 0.68 0.98 0.30 62 77 G 100.8 Satt199 1 Best * 1.61 0.45 0.89 0.4481 77 G 103.2 Satt517 4 * 1.59 0.07 0.51 0.44 58 79 G 137.0 Satt191 2 *1.39 0.18 0.63 0.44 80 79 G 138.4 Sat_117 1 Best * 1.89 0.23 0.77 0.5477 83 H 77.3 Satt279 6 NEW Best * 2.02 0.40 0.90 0.50 63 85 H 125.3Satt181 5 Best ** 3.06 0.05 0.72 0.66 76 86 I 15.5 Satt127 2 Best **2.36 0.64 1.00 0.36 77 87 I 57.9 Satt270 12 NEW Best ** 2.28 0.00 0.040.04† 82 88 I 77.4 Satt292 3 NEW Best * 1.84 0.06 0.56 0.50 64 90 J 19.6SAG1223 4 NEW Best ** 2.59 0.22 0.89 0.67 27 90 J 26.4 SAC1699 3 NEW **1.96 0.04 0.62 0.58 74 91.5 J 61.3 Sat_065 2 Best ** 4.00 0.23 0.96 0.7365 91.5 J 63.6 Satt596 4 * 1.77 0.24 0.80 0.56 64 92 J 68.0 Satt406 2 **3.05 0.19 0.89 0.70 77 92 J 70.8 Satt380 3 ** 2.80 0.29 0.91 0.63 69 92J 72.6 Satt183 1 ** 2.52 0.43 0.96 0.53 84 92 J 74.4 Satt529 1 Best **3.22 0.43 0.99 0.56 73 94 K 17.9 Satt242 4 Best ** 3.05 0.07 0.71 0.6465 97 K 83.7 Satt617 6 NEW Best * 1.68 0.08 0.60 0.53 24 97 K 85.8Satt240 2 * 1.57 0.05 0.60 0.45 74 100 L 15.6 SAT_301 5 NEW Best * 1.970.22 0.83 0.61 78 101 L 33.9 Satt418 2 NEW * 1.44 0.56 0.94 0.37 63 101L 34.6 Satt398 4 Best ** 3.22 0.34 0.94 0.61 71 102 L 42.3 Satt497 3 NEWBest * 1.66 0.60 0.96 0.36 78 103 L 77.1 Satt166 3 Best ** 3.22 0.110.86 0.75 80 103 L 78.0 Satt448 4 * 1.80 0.22 0.74 0.52 65 104 L 118.0Satt373 3 Best ** 4.00 0.02 0.44 0.42 79 104 L 118.6 Satt513 5 ** 2.510.02 0.24 0.22† 64 104 L 124.5 P12394A-1 2 ** 2.64 0.68 1.00 0.32 85 107M 84.2 SAG1048 2 NEW Best ** 4.00 0.01 0.69 0.68 76 108 M 87.6 Satt536 4** 2.12 0.12 0.69 0.58 83 108 M 91.1 Satt175 5 * 1.76 0.18 0.75 0.57 78108 M 98.1 Satt677 2 NEW * 1.34 0.56 0.96 0.40 28 108 M 100.5 Satt680 2NEW Best ** 3.22 0.13 0.86 0.73 71 109 M 115.0 P10615A-1 1 Best * 1.960.09 0.58 0.50 85 109 M 127.3 Satt551 3 NEW * 1.62 0.04 0.49 0.45 59 111M 180.5 SAT_330-DB 1 NEW Best ** 3.00 0.16 0.85 0.69 61 112 N 20.0P13069A-1 3 Best ** 4.00 0.00 0.44 0.44 84 112 N 25.0 P5467A-1 2 NEW **4.00 0.00 0.45 0.45 83 112 N 25.0 P5467A-2 1 * 1.72 0.21 0.63 0.42 86113 N 38.3 SAT_084 2 NEW * 1.37 0.13 0.55 0.42 68 113 N 40.2 SAT_275-DB2 NEW Best * 1.89 0.17 0.73 0.55 62 115 N 83.4 Satt660 2 NEW ** 2.340.10 0.77 0.67 28 115 N 88.6 Satt339 5 Best ** 2.44 0.17 0.77 0.60 80118 O 2.4 Satt358 2 ** 2.35 0.03 0.44 0.41 75 118 O 6.3 Satt487 1 **2.33 0.00 0.02 0.02† 84 118 O 6.3 Satt487 4 Best ** 2.30 0.37 0.89 0.5284 120 O 60.5 Satt420 2 Best * 1.95 0.33 0.85 0.52 84 120 O 65.1 Satt5764 * 1.49 0.20 0.74 0.54 54 120 O 68.9 Satt633 1 NEW * 1.40 0.25 0.720.47 27 123 O 125.0 Satt581 2 Best ** 2.31 0.35 0.89 0.54 85 124 O 153.3Satt153 3 Best ** 2.75 0.49 0.96 0.47 84 124 O 155.1 Satt243 1 * 1.820.61 0.97 0.36 68 UM UM P10793A-1 2 * 1.40 0.10 0.52 0.43 86 UM UMP13560A-1 1 * 1.42 0.22 0.60 0.38 86 UM UM P13561A-1 2 ** 2.51 0.64 1.000.36 82 UM UM S60021-TB 1 NEW * 1.32 0.53 0.95 0.41 28 UM UM S60048-TB 2NEW ** 2.00 0.60 1.00 0.40 28 UM UM S60076-TB 1 NEW * 1.35 0.59 0.930.34 28 UM UM S60148-TB 2 NEW ** 2.22 0.52 0.96 0.45 28 UM UM S60149-TB1 NEW * 1.46 0.36 0.86 0.50 28 UM UM S60201-TB 1 NEW ** 2.35 0.24 0.890.65 28 UM UM S60243-TB 3 NEW ** 2.92 0.27 0.93 0.66 28 UM UM S60326-TB2 NEW ** 2.11 0.43 0.96 0.54 27 UM UM S60338-TB 1 NEW ** 2.60 0.51 1.000.49 25 UM UM S60350-TB 3 NEW ** 2.64 0.08 0.75 0.67 26 UM UM S60361-TB2 NEW ** 3.22 0.14 0.88 0.74 25 UM UM S60422-TB 1 NEW ** 2.17 0.52 1.000.48 27 UM UM S60440-TB 2 NEW * 1.63 0.06 0.59 0.53 28 UM UM S60446-TB 2NEW ** 2.80 0.51 1.00 0.49 28 UM UM S60505-TB 1 NEW ** 2.03 0.51 1.000.49 26 UM UM S60513-TB 2 NEW * 1.70 0.52 0.96 0.45 28 UM UM S60519-TB 2NEW ** 3022 0.44 1.00 0.56 28 UM UM S60536-TB 1 NEW ** 2.64 0.43 1.000.57 28 UM UM S60552-TB 2 NEW * 1.69 0.31 0.88 0.56 28 UM UM S60585-TB 4NEW * 1.45 0.56 0.96 0.40 27 UM UM S60630-TB 3 NEW * 1.48 0.32 0.89 0.5726 UM UM S60728-TB 3 NEW * 1.46 0.45 0.91 0.46 28 UM UM S60812-TB 1NEW * 1.51 0.40 0.87 0.47 27 UM UM SAC1677 3 ** 2.85 0.16 0.82 0.66 84UM UM SAC1724 3 NEW * 1.49 .64 1.00 0.36 27 UM UM SAG1055 5 NEW * 1.620.65 1.00 0.35 28 UM UM Satt111 3 ** 4.00 .40 0.90 .80 83 UM UM Satt2194 NEW * 1.42 0.05 0.42 0.37 80 UM UM Satt299 10 NEW * 1.38 0.65 0.350.30 79 * 95% significance level ** 99% significance level †LOD greaterthan 2.0, increase in frequency less than 25%.

TABLE 13 Favorable Alleles in A3127 Probability of Parent 1 Parent 2inheriting “+” Chromosome Position Allele WILLIAMS ESSEX Progeny A3127allele A1 19.1 1 − + + 0.5 A1 27.1 3 + − + 0.5 A2 0.0 1 − + + 0.5 A2184.0 4 + − + 0.5 B2 45.0 2 + − + 0.5 C1 99.0 4 − + + 0.5 C2 140.9 5 +− + 0.5 D1a 129.2 5 + − + 0.5 D2 93.8 6 − + 0.5 D2 110.4 1 − + + 0.5 E98.0 1 − + + 0.5 F 1.7 3 + − + 0.5 F 85.0 1 + − + 0.5 G 100.8 1 − + +0.5 H 125.3 5 + − + 0.5 J 68.0 2 not segregating K 17.9 4 + − + 0.5 L34.6 4 not segregating L 77.1 3 + − + 0.5 L 124.5 2 not segregating M87.6 4 − + + 0.5 N 20.0 3 not segregating O 6.3 4 not segregating O 37.74 not segregating O 60.5 2 not segregating O 125.0 2 + − + 0.5 O 153.33 + − + 0.5 UM # 2 + − + 0.5 UM # 3 − + + 0.5 UM # 3 − + + 0.5 # of “+”13 10 all 23 0.000000119 alleles → chance of this happening by chance =1 in 8,388,608

TABLE 14 # of SATT591 SATT429 SATT144 SATT270 SATT529 SATT166 sublinesElite line segregating alleles at the above marker loci 91B91 1 and 2 3and 5 4 92M70 1 and 2 2 and 5  4 92B05 3 and 4 2 93B01 1 and 3 2 93M80 5and 12 2 93M90 1 and 3 2

TABLE 15 Isoline seed yield means from field tests Statistical grouping# of plants Mean Yield Genotype (allele) bulked to Seed advantage ofOriginal Subline at indicated marker form # of reps Yield better sublineElite Line Name locus Isoline (locations) (bu/ac) (bu/ac and %) SATT529SATT166 LSD (0.05) = 2.30 91B91 91B91-13 1 3 42 7 37.5 A 91B91 91B91-151 5 24 7 38.0 A 91B91 91B91-23 2 3 14 7 38.8 A 91B91 91B91-25 2 5 7 737.2 A SATT144 SATT270 LSD (0.05) = 3.48 92M70 92M70-12 1 2 14 10 43.4 A92M70 92M70-15 1 5 46 10 44.4 A 92M70 92M70-22 2 2 13 10 43.0 A 92M7092M70-25 2 5 15 10 43.8 A SATT429 LSD (0.05) = 2.01 92B05 92B05-3 3 1755 37.0 A 92B05 92B05-4 4 116 5 37.0 A SATT591 LSD (0.05) = 1.71 93B0193B01-1 1 51 13 38.0 B 93B01 93B01-3 3 28 13 40.1 A 2.1 bu = 5.4%SATT270 LSD (0.05) = 2.54 93M80 93M80-12 12  5 14 48.5 B 93M80 93M80-5 54 14 51.3 A 2.8 bu = 5.6% SATT591 LSD (0.05) = 2.16 93M90 93M90-1 1 3814 50.2 A 3.0 bu = 6.2% 93M90 93M90-3 3 47 14 47.2 B

APPENDIX I Allele Definition Table Marker Allele Size Range bp Sac1006 191.54 92.48 Sac1006 2 93.67 94.41 Sac1006 3 105.49 106.22 Sac1677 1191.96 192.46 Sac1677 2 193.94 194.93 Sac1677 3 205.90 206.91 Sac1677 4332.86 333.35 Sac1677 5 190.20 190.40 Sac1677 6 204.40 204.60 Sat_084 1151.69 152.30 Sat_084 2 153.28 154.30 Sat_084 3 157.85 158.85 Sat_084 4163.84 164.82 Sat_084 5 172.09 172.56 Sat_084 6 160.26 160.46 Sat_090 1336.52 337.36 Sat_090 2 349.84 350.92 Sat_090 3 355.55 356.29 Sat_090 4338.64 339.23 Sat_090 5 340.79 340.09 Sat_090 6 348.04 348.57 Sat_090 7351.54 352.37 Sat_090 8 353.79 354.12 Sat_090 9 357.22 357.86 Sat_104 1266.36 267.50 Sat_104 2 284.72 285.73 Sat_110 1 165.79 166.96 Sat_110 2179.85 180.32 Sat_110 3 181.88 183.02 Sat_110 4 183.72 184.58 Sat_110 5185.65 186.71 Sat_110 6 187.88 188.05 Sat_117 1 210.34 211.45 Sat_117 2218.15 219.41 Sat_117 3 220.13 221.05 Sat_117 4 226.19 226.96 Sat_117 5232.64 232.99 Sat_117 6 212.76 213.12 Sat_117 7 216.86 217.25 Sat_117 8222.62 222.99 Sat_117 9 224.83 224.93 Sat_117 10 242.75 242.85 Sat_11711 252.02 252.23 Sat_117 12 255.97 256.17 Sat_117 13 203.20 203.35Sat_117 14 208.93 209.13 Sat_117 15 234.70 234.90 Satt020 1 147.56148.62 Satt020 2 160.10 160.87 Satt040 1 317.25 318.26 Satt040 2 323.30324.31 Satt040 3 326.83 327.81 Satt040 4 329.75 330.66 Satt040 5 332.71333.35 Satt040 6 320.28 321.05 Satt040 7 335.83 336.33 Satt040 8 308.03308.53 Satt040 9 338.89 339.09 Satt040 10 314.39 314.69 Satt042 1 148.26149.12 Satt042 2 157.59 158.45 Satt042 3 160.69 161.67 Satt042 4 163.71164.64 Satt042 5 154.82 155.22 Satt042 6 160.26 160.46 Satt042 7 166.96167.56 Satt042 8 135.78 135.98 Satt042 9 145.41 145.61 Satt050 1 221.60222.79 Satt050 2 224.66 225.89 Satt050 3 239.80 240.68 Satt050 4 252.17253.84 Satt050 5 255.27 255.57 Satt066 1 160.87 161.87 Satt066 2 105.74106.69 Satt066 3 166.76 167.07 Satt066 4 169.95 171.12 Satt066 5 110.96112.11 Satt066 6 151.75 152.78 Satt066 7 155.32 155.84 Satt066 8 158.20158.86 Satt066 9 164.34 164.82 Satt066 10 168.42 169.19 Satt066 11173.11 174.03 Satt066 12 176.16 176.93 Satt066 13 148.64 149.67 Satt06614 117.63 118.13 Satt092 1 340.69 341.13 Satt092 2 349.07 350.92 Satt0923 332.36 333.35 Satt092 4 338.79 339.25 Satt092 5 341.43 342.56 Satt0926 353.17 353.29 Satt102 1 160.36 161.37 Satt102 2 175.43 176.43 Satt1023 172.56 173.53 Satt102 4 178.87 179.35 Satt102 5 181.79 182.25 Satt1026 184.58 185.65 Satt108 1 169.96 171.12 Satt108 2 173.06 174.00 Satt1083 175.96 177.00 Satt108 4 167.15 167.92 Satt108 5 161.77 161.97 Satt1086 149.12 149.22 Satt109 1 146.99 148.06 Satt109 2 164.62 165.54 Satt1093 167.65 168.42 Satt109 4 162.27 162.47 Satt109 5 144.05 144.33 Satt1111 254.76 255.57 Satt111 2 257.79 258.83 Satt111 3 260.76 261.78 Satt1114 263.82 264.54 Satt111 5 278.98 280.09 Satt111 6 285.22 285.73 Satt1117 288.24 288.75 Satt111 8 266.88 267.90 Satt111 9 270.13 270.33 Satt11110 276.28 276.85 Satt111 11 248.80 249.00 Satt115 1 218.67 219.88Satt115 2 236.35 237.83 Satt115 3 239.29 240.78 Satt115 4 242.65 243.20Satt115 5 221.82 222.22 Satt115 6 238.31 238.51 Satt122 1 234.40 235.37Satt122 2 282.65 283.26 Satt122 3 291.75 292.40 Satt127 1 241.61 242.25Satt127 2 244.22 245.31 Satt127 3 232.44 233.10 Satt127 4 238.81 239.29Satt127 5 247.60 248.20 Satt127 6 250.71 251.22 Satt129 1 120.97 122.06Satt129 2 133.16 134.38 Satt129 3 136.28 137.33 Satt129 4 149.22 150.17Satt129 5 152.23 153.28 Satt129 6 155.84 156.34 Satt129 8 127.42 128.08Satt129 9 140.02 140.52 Satt129 10 143.25 143.75 Satt129 11 132.05132.25 Satt130 1 142.17 143.25 Satt130 2 147.44 148.62 Satt130 3 150.62151.75 Satt130 4 154.30 154.82 Satt131 1 157.85 159.25 Satt131 2 173.06174.03 Satt131 3 175.96 176.93 Satt131 4 179.35 179.85 Satt131 5 182.16182.75 Satt131 6 148.62 149.67 Satt133 1 163.80 164.82 Satt133 2 181.99182.85 Satt133 3 167.36 167.56 Satt133 4 176.06 176.26 Satt133 5 172.96173.41 Satt138 1 226.66 227.82 Satt138 2 229.84 230.75 Satt138 3 232.85233.82 Satt138 4 235.30 236.56 Satt138 5 241.66 242.45 Satt138 6 231.47231.87 Satt138 7 238.91 239.11 Satt138 8 214.97 215.17 Satt142 1 122.55123.25 Satt142 2 137.89 138.69 Satt142 3 141.10 142.17 Satt142 4 144.33145.41 Satt142 5 156.34 156.94 Satt142 6 148.06 148.16 Satt142 7 108.04108.20 Satt142 8 129.00 129.20 Satt144 1 210.72 212.56 Satt144 2 227.62228.58 Satt144 3 231.25 231.40 Satt146 1 291.11 292.36 Satt146 2 294.26295.32 Satt146 3 309.55 310.57 Satt146 4 313.02 314.06 Satt146 5 316.11317.50 Satt146 7 306.71 307.71 Satt146 8 298.10 298.30 Satt146 9 319.97320.38 Satt146 10 322.80 323.30 Satt146 11 329.37 329.87 Satt146 12331.75 332.66 Satt146 13 334.84 335.83 Satt147 1 176.73 177.70 Satt147 2182.75 183.52 Satt147 3 193.45 195.03 Satt147 4 179.95 180.52 Satt147 5207.01 208.72 Satt147 6 211.35 211.55 Satt147 7 213.90 214.77 Satt147 8216.95 217.55 Satt147 9 199.29 199.49 Satt150 1 246.11 248.00 Satt150 2249.20 249.70 Satt150 3 267.70 267.90 Satt150 4 270.95 271.05 Satt150 5282.55 283.21 Satt150 6 234.18 234.60 Satt150 7 237.33 237.63 Satt151 1169.04 170.15 Satt151 2 178.06 179.35 Satt151 3 184.04 185.15 Satt151 4187.08 188.05 Satt151 5 205.40 206.40 Satt151 6 166.29 166.76 Satt151 7175.46 175.96 Satt151 8 198.89 199.39 Satt153 1 198.39 199.49 Satt153 2201.39 202.09 Satt153 3 213.47 214.57 Satt153 4 231.30 232.17 Satt153 5220.13 220.53 Satt155 1 151.75 152.78 Satt155 2 160.87 162.37 Satt155 3163.84 165.02 Satt155 4 167.11 167.72 Satt155 5 158.35 158.45 Satt155 6148.82 149.12 Satt155 7 197.50 197.90 Satt156 1 326.12 326.84 Satt156 2329.20 330.02 Satt156 3 343.89 344.85 Satt156 4 349.65 350.00 Satt156 5346.68 347.11 Satt159 1 268.90 269.53 Satt159 2 269.73 270.33 Satt159 3271.94 272.78 Satt159 4 287.23 287.74 Satt159 5 290.25 290.95 Satt165 1210.44 211.45 Satt165 2 213.37 214.63 Satt165 3 219.41 220.48 Satt165 4216.55 216.95 Satt165 5 204.70 204.90 Satt165 6 225.69 225.99 Satt166 1187.08 188.10 Satt166 2 220.48 221.35 Satt166 3 226.44 227.62 Satt166 4229.48 230.24 Satt166 5 232.37 233.67 Satt166 6 223.39 223.76 Satt166 7217.35 217.75 Satt168 1 345.94 346.71 Satt168 2 359.98 360.77 Satt168 3362.95 363.65 Satt168 4 365.87 367.16 Satt168 5 368.83 369.34 Satt168 6335.26 336.00 Satt168 7 311.10 311.30 Satt168 8 329.57 329.77 Satt172 1155.84 156.89 Satt172 2 167.72 168.52 Satt172 3 170.62 171.52 Satt172 4179.35 180.32 Satt172 5 182.25 183.22 Satt175 1 160.21 161.37 Satt175 2163.31 164.54 Satt175 3 166.29 166.96 Satt175 4 169.19 170.50 Satt175 5178.30 179.55 Satt175 6 193.45 194.74 Satt175 7 172.76 173.26 Satt175 8175.61 176.26 Satt175 9 188.05 188.60 Satt175 10 190.98 191.96 Satt17511 181.69 181.89 Satt176 1 138.39 139.15 Satt176 2 141.60 142.77 Satt1763 160.36 162.37 Satt176 4 169.68 170.77 Satt176 5 172.49 173.68 Satt1766 187.65 188.85 Satt176 7 148.16 149.02 Satt176 8 163.55 164.67 Satt1769 175.56 176.63 Satt176 10 181.62 182.75 Satt181 1 324.31 325.37 Satt1812 346.28 347.16 Satt181 3 354.62 355.60 Satt181 4 357.67 358.46 Satt1815 363.01 364.30 Satt181 6 351.94 352.47 Satt181 7 360.53 361.17 Satt1818 366.01 367.16 Satt181 9 349.07 349.55 Satt183 1 122.04 123.05 Satt1832 128.08 129.10 Satt183 3 144.83 145.91 Satt183 4 148.16 148.62 Satt1835 131.43 131.85 Satt183 6 135.26 135.78 Satt183 7 138.49 138.95 Satt1838 141.70 141.90 Satt186 1 361.81 363.07 Satt186 2 364.72 365.94 Satt1863 367.87 368.83 Satt186 4 339.25 341.09 Satt186 5 359.30 359.73 Satt1866 371.22 371.72 Satt186 7 373.74 374.61 Satt186 8 376.96 377.46 Satt1869 353.39 353.59 Satt190 1 187.01 188.06 Satt190 2 193.35 194.10 Satt1903 196.42 197.40 Satt190 4 225.99 227.06 Satt190 5 181.17 182.03 Satt1906 184.18 184.68 Satt190 7 190.50 190.98 Satt190 8 220.38 220.85 Satt1909 214.07 214.57 Satt191 1 319.67 320.28 Satt191 2 334.84 336.02 Satt1913 337.81 339.02 Satt191 4 349.07 350.00 Satt191 5 352.14 352.77 Satt1916 354.62 355.65 Satt191 7 360.69 361.17 Satt191 8 346.68 346.71 Satt1931 165.79 166.76 Satt193 2 168.94 169.65 Satt193 3 174.59 175.96 Satt1934 180.77 181.79 Satt193 5 186.61 187.75 Satt193 6 189.97 190.60 Satt1937 192.95 193.75 Satt193 8 198.89 199.89 Satt193 9 202.19 203.40 Satt19310 183.72 185.15 Satt193 11 171.79 173.06 Satt193 12 160.11 161.37Satt193 13 177.85 179.08 Satt193 14 163.36 163.84 Satt195 1 214.77216.11 Satt195 2 217.93 218.91 Satt195 3 221.65 222.22 Satt196 1 230.04230.50 Satt196 2 232.94 233.52 Satt196 3 239.29 239.79 Satt196 4 241.76243.00 Satt196 5 253.84 255.27 Satt196 6 236.17 236.45 Satt196 7 257.51257.81 Satt196 8 263.52 263.92 Satt197 1 177.50 179.35 Satt197 2 184.18185.15 Satt197 3 187.08 188.55 Satt197 4 190.00 191.48 Satt197 5 192.95194.44 Satt197 6 205.65 206.00 Satt197 7 170.35 170.72 Satt197 8 175.44175.96 Satt197 9 166.96 167.71 Satt197 10 181.59 181.79 Satt199 1 161.84162.86 Satt199 2 202.22 204.40 Satt199 3 166.29 166.76 Satt199 4 168.22168.69 Satt199 5 174.03 174.99 Satt199 6 178.37 178.87 Satt199 7 220.85221.35 Satt199 8 176.01 176.16 Satt199 9 170.20 170.60 Satt202 1 306.48307.51 Satt202 2 309.55 310.72 Satt202 3 287.33 287.79 Satt202 4 313.03313.66 Satt202 5 285.22 285.93 Satt203 1 223.29 224.06 Satt203 2 229.53230.04 Satt203 3 249.20 250.00 Satt203 4 258.33 259.34 Satt203 5 205.40205.90 Satt203 6 212.56 212.76 Satt203 7 219.61 219.81 Satt203 8 226.51227.82 Satt203 9 231.30 231.67 Satt204 1 234.88 235.87 Satt204 2 240.78241.76 Satt204 3 247.00 248.20 Satt204 4 250.00 250.76 Satt204 5 253.03253.79 Satt204 6 238.13 238.31 Satt204 7 256.59 256.79 Satt209 1 311.80312.63 Satt209 2 329.37 330.48 Satt209 3 310.77 311.10 Satt212 1 347.13348.73 Satt212 2 355.42 356.49 Satt212 3 358.66 358.80 Satt213 1 326.34327.35 Satt213 2 318.46 318.66 Satt216 1 195.43 196.42 Satt216 2 198.79199.39 Satt216 3 222.79 223.29 Satt216 4 225.69 226.66 Satt216 5 210.95211.45 Satt216 6 216.95 217.45 Satt216 7 220.18 220.38 Satt218 1 341.13342.16 Satt218 2 344.12 344.85 Satt219 1 126.38 127.07 Satt219 2 141.95142.67 Satt219 3 157.69 158.55 Satt219 4 175.46 176.58 Satt219 5 120.38120.77 Satt219 6 154.82 155.12 Satt219 7 160.97 161.37 Satt219 8 172.80173.46 Satt219 9 178.78 179.55 Satt219 10 182.09 182.45 Satt220 1 337.71338.29 Satt220 2 340.39 341.23 Satt220 3 343.24 344.42 Satt220 4 346.28346.71 Satt220 5 334.54 334.94 Satt220 6 354.62 355.02 Satt220 7 360.23360.63 Satt220 8 352.52 352.57 Satt221 1 228.12 229.08 Satt221 2 234.12234.90 Satt221 3 237.33 238.31 Satt221 4 246.50 247.50 Satt221 5 231.20231.47 Satt225 1 99.95 101.41 Satt225 2 102.87 103.93 Satt225 3 109.05110.01 Satt227 1 327.28 328.73 Satt227 2 333.35 334.55 Satt228 1 293.26294.26 Satt228 2 298.30 299.06 Satt228 3 310.00 310.57 Satt228 4 323.40324.31 Satt228 5 326.44 327.35 Satt228 6 329.61 330.88 Satt228 7 317.45317.65 Satt228 8 341.53 341.73 Satt230 1 226.14 227.26 Satt230 2 229.33230.50 Satt230 3 231.97 232.99 Satt230 4 214.97 215.47 Satt230 5 223.39224.26 Satt233 1 90.83 91.34 Satt233 4 91.64 92.40 Satt233 5 97.75 98.46Satt233 6 103.73 105.25 Satt233 7 112.77 113.66 Satt234 1 175.32 176.21Satt234 2 178.35 179.35 Satt234 3 172.96 173.06 Satt236 1 250.00 250.81Satt236 2 255.77 256.79 Satt236 3 259.08 259.84 Satt236 4 262.16 262.95Satt236 5 265.46 265.66 Satt236 6 271.35 271.56 Satt236 7 253.39 253.84Satt240 1 310.07 311.30 Satt240 2 313.16 314.63 Satt240 3 316.44 317.86Satt240 4 319.60 320.48 Satt240 5 326.34 326.54 Satt242 1 182.25 183.22Satt242 2 191.48 192.46 Satt242 3 197.40 198.39 Satt242 4 200.39 201.52Satt242 5 203.25 204.40 Satt242 6 209.43 210.44 Satt242 7 212.91 213.47Satt242 8 167.72 168.22 Satt242 9 179.85 180.32 Satt242 10 188.55 189.02Satt242 11 194.93 195.43 Satt242 12 206.91 207.41 Satt242 13 185.65186.11 Satt242 14 171.94 172.29 Satt242 15 175.09 175.34 Satt242 16181.17 181.22 Satt242 17 196.12 196.27 Satt243 1 334.69 335.83 Satt243 2343.48 344.47 Satt243 3 363.37 364.48 Satt243 4 366.24 366.91 Satt243 5357.96 358.16 Satt243 6 337.76 338.69 Satt243 7 346.58 347.06 Satt243 8349.47 349.60 Satt247 1 363.62 365.08 Satt247 2 369.29 370.70 Satt247 3375.25 376.51 Satt247 4 366.63 367.67 Satt247 5 373.15 373.35 Satt247 6381.11 382.25 Satt247 7 378.30 378.50 Satt247 8 355.25 355.45 Satt249 1217.90 218.73 Satt249 2 220.85 221.65 Satt249 3 251.19 252.23 Satt249 4257.31 258.33 Satt249 5 254.61 255.06 Satt249 6 260.96 261.38 Satt249 7236.75 236.95 Satt250 1 189.02 190.00 Satt250 2 207.16 208.02 Satt250 3228.68 228.98 Satt250 4 213.87 213.97 Satt250 5 219.41 219.61 Satt251 1215.92 216.95 Satt251 2 218.91 219.41 Satt251 3 254.26 255.27 Satt251 4257.81 259.08 Satt251 5 266.48 267.50 Satt251 6 209.77 210.95 Satt251 7260.86 261.38 Satt251 8 212.97 213.97 Satt251 9 272.98 273.18 Satt255 1236.85 237.43 Satt255 2 242.75 243.33 Satt255 3 245.71 246.70 Satt255 4248.70 249.30 Satt256 1 338.99 340.19 Satt256 2 348.27 349.50 Satt256 3351.28 352.27 Satt257 1 255.20 256.29 Satt257 2 267.33 268.52 Satt257 3242.25 243.15 Satt257 4 249.20 250.41 Satt257 5 258.48 259.74 Satt257 6261.78 262.18 Satt258 1 156.62 157.69 Satt258 2 159.71 160.71 Satt258 3161.37 161.87 Satt258 4 162.82 163.84 Satt258 5 165.79 166.76 Satt258 6168.22 168.42 Satt259 1 187.08 188.15 Satt259 2 193.45 194.44 Satt259 3205.40 206.50 Satt259 4 207.65 209.58 Satt259 5 211.82 212.64 Satt262 1231.97 232.94 Satt262 2 244.38 245.21 Satt262 3 247.20 248.20 Satt262 4250.21 251.22 Satt262 5 256.29 257.31 Satt262 6 241.41 242.06 Satt262 7253.42 254.04 Satt262 8 235.67 235.87 Satt263 1 249.53 250.21 Satt263 2252.43 253.44 Satt263 3 267.70 268.72 Satt263 4 276.75 278.02 Satt263 5289.18 290.10 Satt263 6 255.72 256.29 Satt263 7 265.04 265.46 Satt263 8271.56 271.76 Satt263 9 246.44 247.20 Satt264 1 216.38 217.45 Satt264 2219.35 220.38 Satt264 3 228.41 229.54 Satt264 4 237.33 238.46 Satt264 5246.65 247.70 Satt264 6 243.63 244.72 Satt265 1 264.64 265.96 Satt265 2267.88 268.52 Satt265 3 285.98 286.93 Satt265 4 289.35 289.55 Satt265 5258.93 259.54 Satt265 6 262.18 262.40 Satt265 7 283.11 283.41 Satt266 1321.79 323.05 Satt266 2 330.98 332.32 Satt266 3 303.20 303.93 Satt267 1232.44 233.42 Satt267 2 241.46 242.25 Satt267 3 250.61 251.32 Satt267 4244.82 245.21 Satt267 5 247.80 248.00 Satt270 1 110.46 110.96 Satt270 2119.10 119.98 Satt270 3 140.97 141.60 Satt270 4 141.86 142.67 Satt270 5144.05 144.98 Satt270 6 150.65 151.28 Satt270 7 116.20 116.71 Satt270 8125.06 125.56 Satt270 9 128.60 129.10 Satt270 10 147.56 148.06 Satt27011 154.31 155.32 Satt270 12 156.61 157.34 Satt272 1 223.76 225.23Satt272 2 229.94 231.00 Satt272 3 226.94 228.12 Satt272 4 233.42 233.92Satt272 5 242.25 242.75 Satt272 6 247.20 247.70 Satt274 1 195.33 196.32Satt274 2 198.38 199.44 Satt274 3 201.34 202.44 Satt274 4 204.40 205.40Satt274 5 180.25 180.52 Satt274 6 189.42 189.62 Satt274 7 207.45 208.72Satt279 1 174.83 175.76 Satt280 1 230.04 231.05 Satt280 2 180.92 181.79Satt280 2 233.19 233.92 Satt280 3 236.17 236.90 Satt280 4 202.69 203.00Satt280 5 224.29 224.98 Satt282 1 314.04 314.79 Satt282 2 317.12 318.31Satt282 3 326.34 327.85 Satt282 4 192.95 193.65 Satt282 4 332.51 334.05Satt282 5 335.68 336.92 Satt282 6 347.84 348.04 Satt282 7 323.45 324.31Satt282 8 329.67 330.58 Satt282 9 338.88 339.75 Satt282 10 310.67 311.70Satt284 1 115.70 116.66 Satt284 2 124.56 125.61 Satt284 3 127.95 128.70Satt284 4 119.20 119.30 Satt284 5 112.86 113.06 Satt284 6 198.89 199.89Satt284 6 129.40 129.61 Satt285 1 208.93 210.19 Satt285 2 231.97 232.95Satt285 3 235.51 236.36 Satt285 4 241.26 242.50 Satt285 5 243.73 245.41Satt285 6 206.40 206.80 Satt285 7 187.28 187.48 Satt285 7 229.32 230.34Satt287 1 203.38 204.40 Satt287 2 233.55 234.40 Satt287 3 197.70 198.09Satt287 4 243.15 243.53 Satt287 9 205.60 205.70 Satt292 1 221.35 221.82Satt292 2 224.26 225.23 Satt292 3 239.29 240.28 Satt292 4 242.66 243.23Satt292 5 251.70 252.23 Satt292 6 229.18 229.48 Satt292 7 232.24 232.44Satt292 8 211.25 211.55 Satt292 9 254.76 255.06 Satt292 10 257.91 258.21Satt295 1 220.38 221.25 Satt295 2 223.29 224.26 Satt295 3 226.57 227.16Satt295 4 232.44 233.14 Satt295 5 244.62 245.21 Satt295 6 256.73 257.41Satt295 7 264.94 265.96 Satt295 8 208.82 209.03 Satt295 9 217.15 217.75Satt295 11 235.62 236.02 Satt295 12 238.81 239.01 Satt295 13 241.66241.91 Satt295 14 253.84 254.04 Satt299 1 258.01 258.21 Satt299 2 283.51283.61 Satt299 3 284.01 284.97 Satt299 4 285.12 286.27 Satt299 5 291.25292.30 Satt299 6 303.70 304.30 Satt299 7 306.88 307.51 Satt299 8 313.66314.69 Satt299 9 318.76 319.47 Satt299 10 328.65 329.87 Satt299 11331.87 332.81 Satt299 12 342.56 343.49 Satt299 13 293.26 293.36 Satt3001 238.81 239.79 Satt300 2 245.20 245.81 Satt300 3 254.26 255.27 Satt3004 266.48 267.23 Satt300 5 269.53 270.55 Satt300 6 242.25 242.85 Satt3007 257.81 258.11 Satt300 8 263.57 263.82 Satt303 1 128.60 129.10 Satt3032 131.59 132.15 Satt303 3 134.68 135.34 Satt303 4 137.82 138.39 Satt3035 140.96 141.70 Satt303 6 144.13 144.88 Satt303 7 153.65 154.41 Satt3038 156.67 157.49 Satt303 9 119.59 120.08 Satt303 10 122.75 123.05 Satt30311 147.66 147.86 Satt303 12 159.96 160.16 Satt304 1 98.35 99.16 Satt3042 162.75 163.84 Satt304 3 165.99 166.59 Satt304 4 168.81 169.65 Satt3071 336.32 337.51 Satt307 2 350.42 351.44 Satt307 3 355.99 356.92 Satt3074 358.93 360.82 Satt307 5 362.00 362.67 Satt307 6 365.33 365.84 Satt3077 367.97 368.48 Satt311 1 152.78 153.81 Satt311 2 164.82 165.79 Satt3113 175.96 176.43 Satt311 4 156.24 156.34 Satt311 5 192.37 192.47 Satt3116 207.41 207.51 Satt311 7 201.44 201.89 Satt311 8 210.14 210.85 Satt3119 181.29 181.69 Satt314 1 241.26 242.25 Satt314 2 244.22 245.21 Satt3143 247.50 247.80 Satt319 1 173.31 174.59 Satt319 2 176.48 177.50 Satt3193 179.47 180.52 Satt319 4 218.91 219.88 Satt319 5 237.16 238.13 Satt3196 240.78 241.26 Satt319 7 183.22 183.32 Satt319 8 192.27 192.47 Satt3199 210.44 210.95 Satt319 10 234.90 235.00 Satt321 1 231.97 233.25 Satt3212 235.25 236.23 Satt321 3 238.31 239.26 Satt321 4 244.22 244.72 Satt3221 313.03 313.81 Satt322 2 312.13 312.78 Satt326 1 323.68 324.81 Satt3262 326.69 327.85 Satt326 3 329.87 330.88 Satt326 4 342.06 342.76 Satt3265 320.83 321.39 Satt327 1 251.22 252.23 Satt327 2 254.26 255.28 Satt3273 245.41 245.81 Satt327 4 248.55 248.80 Satt327 5 233.62 234.02 Satt3276 263.72 263.92 Satt327 7 242.42 242.80 Satt327 8 257.51 257.66 Satt3281 248.20 249.20 Satt328 2 251.22 252.23 Satt329 1 238.31 238.81 Satt3292 268.52 269.53 Satt329 3 272.58 273.38 Satt329 4 275.63 276.65 Satt3295 279.69 280.70 Satt329 6 247.20 247.70 Satt329 7 254.26 254.76 Satt3298 257.31 258.33 Satt329 9 267.50 268.00 Satt329 10 277.25 277.45 Satt32911 279.28 279.48 Satt329 12 274.00 274.10 Satt329 13 270.13 270.23Satt330 1 308.03 308.78 Satt330 2 342.41 343.49 Satt330 3 348.04 348.72Satt330 4 355.55 355.99 Satt330 5 364.02 364.78 Satt330 6 344.72 344.85Satt330 7 345.50 345.98 Satt330 8 346.71 346.91 Satt330 9 316.53 316.93Satt331 1 233.42 234.40 Satt331 2 239.67 240.28 Satt331 3 251.56 252.43Satt331 4 236.95 237.15 Satt332 1 161.69 162.37 Satt332 2 176.57 177.90Satt332 3 179.65 180.32 Satt332 4 182.85 183.20 Satt332 5 173.78 174.13Satt333 1 180.32 180.82 Satt333 2 195.43 196.47 Satt333 3 198.89 199.39Satt333 4 201.89 202.39 Satt333 5 204.74 205.40 Satt333 6 171.79 171.99Satt333 7 189.82 190.00 Satt333 8 177.75 178.20 Satt333 9 192.76 193.10Satt334 1 202.39 202.90 Satt334 2 207.92 208.93 Satt334 3 210.95 211.96Satt334 4 214.33 215.77 Satt334 5 215.96 216.76 Satt334 6 217.93 218.70Satt334 7 220.38 221.35 Satt334 8 196.12 196.32 Satt334 9 230.34 230.50Satt335 1 145.31 146.49 Satt335 2 154.82 156.09 Satt335 3 164.34 165.27Satt335 4 167.51 168.22 Satt335 5 170.55 171.27 Satt336 1 176.43 177.40Satt336 2 182.53 183.22 Satt336 3 185.50 186.61 Satt336 4 191.48 192.46Satt338 1 229.08 230.04 Satt338 2 232.54 233.92 Satt338 3 253.24 254.37Satt338 4 265.46 266.68 Satt338 5 274.62 275.57 Satt338 6 277.67 278.68Satt338 7 238.31 238.71 Satt338 8 280.90 281.90 Satt339 1 234.90 235.38Satt339 2 240.47 241.97 Satt339 3 243.73 244.72 Satt339 4 255.91 257.31Satt339 5 264.88 266.26 Satt339 6 268.14 269.07 Satt339 7 206.70 206.91Satt339 8 252.83 253.92 Satt339 9 271.45 271.56 Satt339 10 250.21 250.61Satt339 11 262.28 262.40 Satt339 12 259.13 259.35 Satt343 1 138.39138.95 Satt343 2 141.50 142.37 Satt343 3 160.14 161.87 Satt343 4 169.19169.87 Satt343 5 172.09 173.06 Satt343 6 187.08 188.05 Satt343 7 148.16148.62 Satt343 8 181.12 181.99 Satt343 9 163.26 164.00 Satt343 10 175.46175.86 Satt343 11 154.51 154.71 Satt346 1 321.29 322.30 Satt346 2 327.35328.36 Satt346 3 336.32 337.41 Satt346 4 339.25 340.29 Satt346 5 318.20319.27 Satt346 6 324.40 325.32 Satt347 1 345.25 346.28 Satt347 2 347.94349.07 Satt347 3 351.02 351.84 Satt348 1 308.03 308.72 Satt348 2 333.01333.85 Satt348 3 341.83 342.66 Satt348 4 344.67 345.35 Satt348 5 330.27330.58 Satt348 6 326.84 327.35 Satt348 7 339.55 339.85 Satt352 1 334.54335.83 Satt352 2 341.13 341.63 Satt352 3 346.15 347.21 Satt352 4 351.72352.87 Satt352 5 354.62 355.55 Satt352 6 340.59 340.89 Satt352 7 343.79343.92 Satt352 8 369.49 369.69 Satt352 9 349.17 349.80 Satt352 10 337.41338.54 Satt352 11 360.53 360.63 Satt353 1 144.20 145.03 Satt353 2 159.86160.87 Satt353 3 169.19 169.75 Satt353 4 174.94 175.96 Satt353 5 184.18185.08 Satt353 6 178.37 178.97 Satt353 7 165.84 166.96 Satt353 8 154.30154.50 Satt355 1 248.20 249.20 Satt355 2 257.22 258.33 Satt355 3 272.58273.43 Satt355 4 236.35 237.33 Satt355 5 254.26 254.86 Satt355 6 269.53270.03 Satt355 7 275.88 276.33 Satt355 8 278.88 279.28 Satt355 9 260.76260.96 Satt356 1 322.24 323.30 Satt356 2 325.15 326.72 Satt357 1 171.69173.06 Satt357 2 275.63 277.15 Satt357 3 278.60 280.19 Satt357 4 183.22184.18 Satt357 5 198.70 199.39 Satt357 6 262.40 262.90 Satt357 7 269.53270.03 Satt357 8 286.73 287.23 Satt358 1 194.93 196.42 Satt358 2 206.91208.17 Satt358 3 209.94 210.95 Satt358 4 203.90 205.10 Satt358 5 164.82166.29 Satt358 6 198.39 199.39 Satt358 7 162.85 163.84 Satt359 1 182.58183.65 Satt359 2 184.68 186.00 Satt359 3 197.75 198.64 Satt359 4 200.89201.39 Satt359 5 189.12 189.42 Satt359 6 203.80 204.50 Satt359 7 206.75207.36 Satt359 8 164.82 165.27 Satt361 1 271.81 273.08 Satt361 2 274.95275.93 Satt361 3 269.02 269.93 Satt361 4 278.07 278.78 Satt364 1 232.45233.40 Satt364 2 235.38 236.18 Satt364 3 241.26 242.55 Satt364 4 244.22245.41 Satt364 5 238.61 238.91 Satt364 6 229.54 229.74 Satt367 1 191.96193.00 Satt367 2 212.97 214.07 Satt367 3 215.96 217.15 Satt367 4 221.82223.09 Satt367 5 225.13 226.19 Satt367 6 228.12 229.18 Satt367 7 231.25232.07 Satt367 8 195.58 195.83 Satt367 9 189.07 189.47 Satt367 10 234.70234.80 Satt367 11 196.72 197.10 Satt367 12 210.34 210.64 Satt369 1234.60 236.18 Satt369 2 236.65 237.33 Satt369 3 239.55 240.78 Satt369 4242.51 243.73 Satt369 5 218.43 219.06 Satt369 6 227.53 228.02 Satt369 7231.00 231.20 Satt369 8 245.63 246.11 Satt372 1 336.98 338.29 Satt372 2340.05 341.13 Satt372 3 345.78 346.28 Satt372 4 348.57 349.17 Satt372 5351.34 352.27 Satt372 6 334.15 334.54 Satt372 7 350.52 350.62 Satt372 8306.08 306.58 Satt373 1 143.25 143.75 Satt373 2 146.49 147.09 Satt373 3149.52 150.27 Satt373 4 158.75 159.51 Satt373 5 179.85 180.32 Satt373 6182.70 183.72 Satt373 7 212.97 213.47 Satt373 8 152.78 153.28 Satt373 9155.72 156.54 Satt373 10 173.73 174.43 Satt373 11 178.20 178.58 Satt37312 185.85 186.11 Satt373 13 191.97 192.37 Satt373 14 194.93 195.33Satt373 15 198.10 198.40 Satt373 16 215.96 216.71 Satt373 17 218.80219.66 Satt373 18 207.11 207.31 Satt378 1 138.70 139.45 Satt378 2 151.75152.78 Satt378 3 145.61 145.81 Satt378 4 148.82 149.22 Satt378 5 159.86160.26 Satt380 1 147.94 148.82 Satt380 2 149.22 149.82 Satt380 3 151.31152.15 Satt380 4 152.35 152.78 Satt383 1 341.04 341.89 Satt383 2 345.72346.86 Satt383 3 348.45 349.60 Satt383 4 351.32 352.42 Satt383 5 340.29340.49 Satt383 6 344.22 344.42 Satt384 1 125.06 125.76 Satt384 2 153.68154.45 Satt384 3 156.80 157.34 Satt385 1 100.37 101.41 Satt385 2 115.70116.30 Satt385 3 128.08 128.60 Satt385 4 140.32 141.60 Satt385 5 143.45144.53 Satt385 6 149.91 151.11 Satt385 7 146.97 148.06 Satt385 8 156.29157.34 Satt385 9 160.87 161.37 Satt385 10 163.84 164.34 Satt385 11173.06 174.29 Satt385 12 153.18 153.38 Satt387 1 171.12 171.59 Satt387 2177.90 178.37 Satt387 3 200.35 201.14 Satt387 4 204.90 206.40 Satt387 5212.97 213.97 Satt387 6 216.16 216.36 Satt389 1 145.91 146.49 Satt389 2148.87 149.67 Satt389 3 153.28 154.21 Satt389 4 173.49 174.62 Satt389 5176.43 177.45 Satt389 6 179.45 180.94 Satt389 7 147.96 148.26 Satt389 8156.44 156.84 Satt389 9 182.45 183.15 Satt389 10 140.52 140.82 Satt38911 168.22 168.42 Satt390 1 358.94 359.73 Satt390 2 361.62 362.77 Satt3903 356.49 356.92 Satt390 4 364.73 365.44 Satt390 5 350.67 350.87 Satt3911 168.69 169.65 Satt391 2 183.67 184.68 Satt391 3 181.12 181.29 Satt3914 187.28 187.48 Satt393 1 325.13 326.20 Satt393 2 328.15 329.16 Satt3933 322.09 322.50 Satt393 4 331.47 331.67 Satt398 1 151.61 152.78 Satt3982 154.70 155.84 Satt398 3 157.71 158.86 Satt398 4 166.76 167.92 Satt3985 169.85 171.12 Satt398 6 160.79 161.87 Satt398 7 163.89 164.82 Satt3988 176.43 176.93 Satt398 9 179.85 180.32 Satt398 10 182.25 183.72 Satt39811 148.62 148.82 Satt399 1 312.51 313.76 Satt399 2 325.00 326.12 Satt3993 328.05 329.37 Satt399 4 331.31 332.12 Satt406 1 161.57 162.57 Satt4062 239.21 240.38 Satt406 3 242.25 243.33 Satt406 4 245.41 246.20 Satt4065 158.73 159.46 Satt406 6 233.42 233.82 Satt406 7 230.34 230.44 Satt4068 215.47 215.57 Satt406 9 236.28 237.26 Satt406 10 227.26 227.46 Satt4091 166.29 166.76 Satt409 2 169.19 170.15 Satt409 3 174.99 176.16 Satt4094 184.18 185.20 Satt409 5 187.08 188.35 Satt409 6 193.68 194.44 Satt4097 199.69 200.39 Satt409 8 196.72 196.90 Satt409 9 157.24 157.54 Satt40910 163.56 163.66 Satt409 11 202.79 202.90 Satt409 12 190.51 191.28Satt409 13 172.29 172.39 Satt411 1 97.95 99.06 Satt411 2 100.93 102.21Satt411 3 93.77 94.17 Satt412 1 248.70 249.57 Satt412 2 254.76 255.78Satt412 3 257.79 258.83 Satt412 4 260.81 261.88 Satt412 5 263.87 264.94Satt412 6 251.91 252.73 Satt412 7 267.23 268.00 Satt413 1 333.87 334.84Satt413 2 356.86 357.96 Satt413 3 359.66 360.72 Satt413 4 369.64 370.75Satt413 5 342.16 342.36 Satt413 6 342.96 342.99 Satt413 7 366.61 366.81Satt413 8 368.63 369.49 Satt414 1 277.87 279.18 Satt414 2 317.75 319.16Satt414 3 321.22 322.30 Satt414 4 324.15 325.32 Satt414 5 327.35 328.36Satt414 7 336.62 337.31 Satt414 8 302.69 303.40 Satt414 9 315.21 316.23Satt414 10 339.70 340.19 Satt414 11 342.66 342.86 Satt414 12 330.78331.28 Satt415 1 290.25 291.40 Satt415 2 297.19 298.60 Satt415 3 300.33301.87 Satt415 4 303.75 304.96 Satt415 5 310.07 310.97 Satt415 6 323.10323.50 Satt415 7 292.05 292.35 Satt415 8 307.61 307.96 Satt415 9 288.66289.45 Satt416 1 241.96 242.16 Satt416 2 253.32 254.26 Satt416 3 256.56257.91 Satt416 4 262.64 263.42 Satt416 5 265.86 266.48 Satt416 6 280.82282.30 Satt416 7 244.34 244.77 Satt416 8 259.66 260.05 Satt417 1 299.06300.33 Satt417 2 318.26 319.27 Satt417 3 321.44 322.30 Satt417 4 339.40341.13 Satt417 5 342.44 343.92 Satt417 6 336.96 337.51 Satt418 1 207.40208.42 Satt418 2 210.36 211.45 Satt418 3 213.40 214.47 Satt418 4 216.35217.45 Satt418 5 219.41 219.88 Satt418 6 189.53 190.00 Satt418 7 192.47193.45 Satt418 8 198.60 199.39 Satt418 9 204.50 204.85 Satt420 1 249.67250.71 Satt420 2 260.72 261.97 Satt420 3 266.98 268.00 Satt420 4 274.62275.12 Satt420 5 252.88 253.74 Satt420 6 256.08 256.29 Satt420 7 264.22264.44 Satt421 1 135.14 136.43 Satt421 2 144.83 146.49 Satt421 3 148.55149.22 Satt422 1 311.60 312.77 Satt422 2 321.29 321.79 Satt422 3 330.38331.74 Satt422 4 333.35 334.34 Satt422 5 315.31 315.61 Satt422 6 318.36319.30 Satt422 7 336.92 337.12 Satt422 8 327.85 328.05 Satt423 1 241.76242.75 Satt423 2 262.90 263.92 Satt423 3 266.06 266.98 Satt423 4 340.05340.69 Satt423 6 321.79 321.89 Satt423 7 248.60 248.80 Satt426 1 198.39199.59 Satt426 2 200.39 200.89 Satt426 3 202.39 203.40 Satt426 4 203.90204.40 Satt426 5 205.34 206.40 Satt426 6 217.55 218.03 Satt426 7 192.46192.66 Satt426 8 223.59 223.76 Satt426 9 196.90 197.00 Satt429 1 263.42264.22 Satt429 2 266.48 267.50 Satt429 3 269.53 270.65 Satt429 4 272.54273.65 Satt429 5 245.21 246.11 Satt429 6 248.20 248.85 Satt429 7 254.46254.76 Satt429 8 275.83 276.43 Satt429 9 257.31 257.71 Satt430 1 241.76242.75 Satt430 2 244.72 245.41 Satt430 3 248.10 248.70 Satt430 4 254.26254.46 Satt430 5 257.31 257.51 Satt430 6 239.21 239.30 Satt431 1 305.46306.38 Satt431 2 318.20 318.96 Satt431 3 321.29 322.30 Satt431 4 324.31324.81 Satt431 5 342.51 343.29 Satt431 6 348.11 349.07 Satt431 7 315.41315.51 Satt431 8 331.37 331.57 Satt431 9 328.05 328.25 Satt431 10 340.15340.29 Satt431 11 345.50 345.88 Satt431 12 351.44 351.54 Satt432 1255.78 256.29 Satt432 2 258.53 259.50 Satt432 3 261.72 262.40 Satt432 4264.89 265.46 Satt433 1 125.56 126.50 Satt433 2 134.74 135.78 Satt433 3144.33 145.41 Satt433 4 153.80 154.98 Satt433 5 166.14 167.20 Satt433 6156.64 156.94 Satt433 7 138.19 138.49 Satt433 8 159.55 159.86 Satt436 1187.58 188.75 Satt436 2 208.93 209.94 Satt436 3 244.72 245.76 Satt436 4226.96 227.26 Satt436 5 247.70 248.85 Satt436 6 250.71 251.92 Satt436 7203.30 203.60 Satt436 8 254.11 254.76 Satt440 1 286.23 287.23 Satt440 2307.74 308.88 Satt440 3 320.28 320.98 Satt440 4 338.74 339.58 Satt440 5287.33 288.04 Satt440 6 323.70 324.36 Satt440 7 335.80 336.32 Satt440 8317.45 317.65 Satt441 1 161.80 162.77 Satt441 2 164.82 165.79 Satt441 3173.68 174.69 Satt441 4 176.69 177.70 Satt441 5 179.78 180.62 Satt441 6191.90 192.86 Satt441 7 207.02 207.92 Satt441 8 171.07 171.59 Satt441 10168.42 168.52 Satt441 11 136.72 137.43 Satt441 12 152.88 153.48 Satt44113 158.95 159.75 Satt441 14 155.84 156.44 Satt441 15 195.03 195.70Satt441 16 185.85 186.39 Satt441 17 189.52 189.72 Satt442 1 240.28240.78 Satt442 2 246.20 247.30 Satt442 3 249.20 250.21 Satt442 4 252.23253.24 Satt442 5 261.68 262.40 Satt442 6 264.44 265.46 Satt442 7 258.63259.03 Satt442 8 243.66 244.13 Satt442 9 267.80 268.20 Satt442 10 237.63238.03 Satt442 11 255.72 256.29 Satt442 12 283.01 283.31 Satt442 13270.75 271.00 Satt444 1 134.84 136.28 Satt444 2 137.94 138.95 Satt445 1166.18 166.96 Satt445 2 169.02 169.85 Satt445 3 184.28 185.15 Satt445 4193.35 194.54 Satt445 5 196.52 197.40 Satt445 6 215.17 215.47 Satt445 7163.11 163.76 Satt445 8 190.20 190.80 Satt445 9 178.78 179.08 Satt445 10187.38 187.83 Satt445 11 217.45 217.90 Satt448 1 340.69 341.63 Satt448 2347.47 348.19 Satt448 3 350.42 350.92 Satt448 4 355.86 356.92 Satt448 5361.62 362.12 Satt448 6 358.66 359.34 Satt448 7 364.68 365.08 Satt448 8343.59 343.92 Satt448 9 332.07 332.27 Satt448 10 324.81 325.01 Satt451 1325.32 326.49 Satt451 2 351.84 352.37 Satt451 3 354.42 355.25 Satt451 4357.52 357.72 Satt451 5 319.67 319.92 Satt452 1 256.74 257.81 Satt452 2259.68 260.86 Satt452 3 263.42 263.92 Satt452 4 265.81 266.98 Satt452 5274.98 275.83 Satt452 6 296.79 296.99 Satt452 7 254.14 254.36 Satt452 8302.57 303.40 Satt452 9 272.46 272.78 Satt452 10 309.45 309.55 Satt454 1158.70 159.60 Satt454 2 170.40 171.59 Satt454 3 173.53 174.49 Satt454 4176.63 177.50 Satt454 5 161.87 162.37 Satt454 6 182.95 183.15 Satt455 1273.08 274.10 Satt455 2 276.03 277.15 Satt455 3 279.31 280.19 Satt457 1356.79 357.42 Satt457 2 368.33 369.29 Satt457 3 324.81 325.11 Satt457 4336.67 337.12 Satt457 5 354.12 354.32 Satt457 6 365.44 366.09 Satt457 7371.42 371.82 Satt460 1 113.80 114.50 Satt460 2 116.66 117.63 Satt460 3135.24 136.08 Satt460 4 144.83 145.61 Satt460 5 160.36 161.17 Satt460 6163.46 164.24 Satt460 7 129.40 129.61 Satt461 1 128.60 129.61 Satt461 2131.65 132.67 Satt461 3 138.39 138.95 Satt461 4 144.83 145.41 Satt461 5154.30 154.82 Satt461 6 122.95 124.05 Satt461 7 153.28 153.80 Satt461 8134.94 135.24 Satt464 1 144.77 146.31 Satt464 2 164.65 165.79 Satt464 3174.13 175.55 Satt464 4 168.02 168.42 Satt464 5 177.90 178.10 Satt466 1165.27 166.29 Satt466 2 168.32 169.19 Satt466 3 177.80 178.77 Satt467 1274.79 275.52 Satt467 2 294.84 295.98 Satt469 1 245.81 246.90 Satt469 2276.65 277.20 Satt469 3 279.69 280.24 Satt469 4 225.63 225.89 Satt469 5249.50 249.80 Satt470 1 263.62 264.94 Satt470 2 266.48 268.00 Satt471 1233.92 234.60 Satt471 2 245.71 246.70 Satt471 3 248.70 249.70 Satt473 1307.51 308.53 Satt473 2 310.57 311.60 Satt473 3 313.86 314.69 Satt473 4317.25 317.75 Satt473 5 295.27 295.78 Satt473 6 298.30 298.81 Satt473 7301.35 302.37 Satt473 8 304.43 305.46 Satt473 9 292.20 292.35 Satt475 1150.47 151.46 Satt475 2 159.86 160.87 Satt475 3 171.92 173.06 Satt475 4175.46 175.66 Satt475 5 178.10 178.30 Satt476 1 343.49 344.02 Satt476 2346.43 347.21 Satt476 3 354.62 355.55 Satt476 4 357.52 358.36 Satt476 5365.94 366.91 Satt476 6 349.17 349.37 Satt476 7 351.94 352.27 Satt476 8360.53 360.67 Satt476 9 363.37 363.72 Satt476 10 325.62 325.92 Satt477 1141.60 142.67 Satt477 2 157.34 158.35 Satt477 3 163.62 164.44 Satt477 4154.71 155.42 Satt477 5 161.07 161.27 Satt478 1 194.36 195.23 Satt478 2227.16 227.82 Satt478 3 232.95 234.12 Satt478 4 236.28 236.86 Satt478 5239.30 240.15 Satt478 6 242.25 243.03 Satt478 7 254.26 255.11 Satt478 8257.31 258.06 Satt478 9 224.41 224.73 Satt478 10 230.40 230.70 Satt47811 251.42 251.82 Satt478 12 248.40 248.75 Satt478 13 221.56 221.66Satt478 14 221.26 221.46 Satt479 1 114.30 115.40 Satt479 2 117.63 118.50Satt479 3 142.52 143.25 Satt479 4 145.41 146.49 Satt479 5 151.75 152.83Satt479 6 111.76 112.01 Satt480 1 137.81 138.95 Satt480 2 140.99 142.17Satt480 3 144.33 145.41 Satt487 1 114.30 114.90 Satt487 2 117.16 118.13Satt487 3 120.28 121.07 Satt487 4 123.05 124.20 Satt487 5 126.31 127.07Satt487 6 111.36 111.91 Satt487 7 132.67 132.87 Satt487 8 135.78 135.98Satt487 9 129.61 129.91 Satt488 1 295.57 295.78 Satt488 2 311.35 312.33Satt488 3 314.61 315.41 Satt488 4 323.85 324.71 Satt488 5 327.29 327.75Satt491 1 195.86 196.52 Satt491 2 201.78 202.49 Satt491 3 207.41 208.42Satt491 4 216.95 217.35 Satt491 5 219.98 220.28 Satt492 1 235.72 237.13Satt492 2 238.81 239.97 Satt492 3 227.46 227.62 Satt493 1 266.41 267.50Satt493 2 269.83 270.55 Satt493 3 281.93 282.71 Satt493 4 279.23 279.38Satt495 1 231.00 231.97 Satt495 2 243.11 244.22 Satt495 3 237.33 237.83Satt497 1 268.82 269.53 Satt497 2 271.96 272.58 Satt497 3 274.62 276.23Satt497 4 277.67 279.03 Satt497 5 296.18 297.49 Satt497 6 299.31 300.43Satt497 7 281.10 281.90 Satt497 8 265.86 266.06 Satt497 9 262.80 263.20Satt503 1 226.19 227.00 Satt503 2 228.90 230.09 Satt503 3 231.92 233.15Satt503 4 221.83 222.55 Satt503 5 211.15 211.65 Satt503 6 224.93 225.23Satt503 7 234.95 235.59 Satt503 8 223.00 223.10 Satt506 1 280.19 281.35Satt506 2 283.21 284.01 Satt506 3 292.26 293.61 Satt506 4 289.75 290.15Satt507 1 230.60 231.47 Satt507 2 233.42 234.40 Satt507 3 236.36 237.43Satt507 4 227.62 228.02 Satt507 5 239.70 239.90 Satt508 1 333.06 334.15Satt508 2 336.15 337.31 Satt508 3 339.25 339.65 Satt509 1 192.47 193.55Satt509 2 195.43 196.52 Satt509 3 198.40 199.39 Satt509 4 201.39 202.59Satt509 5 204.78 205.40 Satt509 6 236.86 238.41 Satt509 7 249.20 250.41Satt509 8 240.68 241.18 Satt509 9 243.79 244.22 Satt509 10 184.18 184.48Satt509 11 252.83 253.13 Satt510 1 109.05 109.55 Satt510 2 127.07 127.58Satt510 3 136.08 137.33 Satt510 4 139.35 140.52 Satt510 5 96.44 97.45Satt510 6 121.07 122.06 Satt510 7 111.91 112.86 Satt510 8 130.11 130.31Satt511 1 255.83 256.79 Satt511 2 262.02 262.90 Satt511 3 265.03 266.01Satt511 4 268.06 269.07 Satt511 5 277.87 278.07 Satt511 6 259.23 259.55Satt512 1 316.16 317.25 Satt512 2 319.37 320.28 Satt512 3 325.53 326.44Satt512 4 328.71 329.57 Satt512 5 331.77 332.86 Satt512 6 310.07 310.62Satt512 7 313.03 313.68 Satt513 1 117.63 118.13 Satt513 2 130.11 130.63Satt513 3 142.17 143.25 Satt513 4 173.06 173.83 Satt513 5 181.79 182.75Satt513 6 185.05 185.65 Satt513 7 170.15 170.62 Satt513 8 164.34 164.44Satt513 9 167.26 167.63 Satt513 10 133.07 133.37 Satt513 11 139.14139.65 Satt513 12 148.77 148.92 Satt513 13 153.38 153.58 Satt513 14145.81 146.11 Satt514 1 314.34 315.21 Satt514 2 316.63 317.25 Satt514 3325.76 326.34 Satt514 4 351.18 351.94 Satt514 5 328.96 329.26 Satt514 6329.77 330.38 Satt514 7 331.97 332.36 Satt514 8 337.12 337.71 Satt514 9340.15 340.39 Satt514 10 328.25 328.56 Satt514 11 345.78 346.18 Satt51412 359.93 360.13 Satt514 13 362.87 363.17 Satt514 14 354.22 354.62Satt514 15 334.25 334.34 Satt515 1 341.13 342.06 Satt515 2 352.77 353.69Satt515 3 355.05 356.44 Satt517 1 159.30 160.36 Satt517 2 162.12 163.56Satt517 3 165.13 166.49 Satt517 4 173.15 174.50 Satt517 5 176.21 177.23Satt517 6 179.45 180.15 Satt517 7 149.89 150.91 Satt517 8 170.05 171.02Satt519 1 354.60 355.55 Satt519 2 357.42 358.36 Satt519 3 360.33 361.17Satt519 4 363.27 363.47 Satt519 5 372.02 372.18 Satt522 1 231.47 232.34Satt522 2 240.78 241.26 Satt522 3 255.78 256.89 Satt522 4 258.83 259.90Satt522 5 283.31 284.36 Satt522 6 237.53 238.13 Satt522 7 252.83 253.39Satt522 8 280.29 280.70 Satt523 1 171.58 172.39 Satt523 2 174.43 175.34Satt523 3 189.52 190.40 Satt523 4 192.76 193.45 Satt523 5 184.02 184.18Satt523 6 199.04 199.39 Satt524 1 168.22 168.94 Satt524 2 171.12 172.29Satt524 3 177.40 177.60 Satt524 4 174.59 174.89 Satt526 1 322.80 324.22Satt526 2 332.02 333.44 Satt529 1 214.47 215.47 Satt529 2 220.38 221.35Satt529 3 217.85 218.08 Satt529 4 229.67 230.24 Satt529 5 223.96 224.06Satt532 1 170.15 171.37 Satt532 2 172.86 174.48 Satt532 3 179.35 180.37Satt532 4 182.50 183.32 Satt532 5 154.40 154.82 Satt532 6 175.31 176.55Satt532 7 176.58 177.40 Satt532 8 167.48 168.17 Satt533 1 169.19 170.15Satt533 2 174.99 175.96 Satt533 3 193.94 194.44 Satt533 4 196.72 196.90Satt533 5 172.23 172.96 Satt533 6 178.37 178.57 Satt533 7 193.15 193.55Satt534 1 165.79 166.90 Satt534 2 172.01 173.06 Satt534 3 177.90 178.87Satt534 4 180.82 181.84 Satt534 5 183.72 185.15 Satt534 6 187.08 188.55Satt534 7 190.00 191.28 Satt534 8 192.95 194.44 Satt534 9 195.92 197.05Satt534 10 163.36 163.84 Satt534 11 160.36 160.76 Satt534 12 169.18169.85 Satt534 13 157.34 157.74 Satt534 14 175.04 175.96 Satt534 15199.89 200.19 Satt534 16 150.71 151.11 Satt536 1 220.76 221.83 Satt536 2223.76 224.26 Satt536 3 226.66 227.72 Satt536 4 229.38 230.70 Satt536 5197.20 197.50 Satt536 6 232.95 234.22 Satt536 7 236.18 236.61 Satt537 1292.58 293.76 Satt537 2 295.78 296.79 Satt537 3 311.28 312.63 Satt537 4317.75 318.36 Satt537 5 320.67 321.79 Satt537 6 323.80 324.81 Satt537 7301.35 302.62 Satt537 8 308.03 309.10 Satt537 9 280.43 281.20 Satt537 10289.75 290.25 Satt537 11 299.31 299.83 Satt537 12 305.46 305.98 Satt53713 314.69 315.21 Satt537 14 333.35 333.85 Satt537 15 283.61 283.91Satt540 1 175.14 176.06 Satt540 2 181.17 182.25 Satt540 3 190.28 191.48Satt540 4 192.71 194.19 Satt540 5 199.39 200.39 Satt540 6 210.59 211.96Satt540 7 213.37 213.85 Satt540 8 169.65 170.15 Satt540 9 184.68 185.15Satt540 10 187.58 188.05 Satt540 12 195.73 196.12 Satt540 13 196.56197.20 Satt540 14 204.70 204.90 Satt543 1 171.12 172.39 Satt543 2 174.39175.09 Satt543 3 175.29 176.26 Satt543 4 195.43 196.52 Satt543 5 198.83199.49 Satt543 6 207.41 208.42 Satt543 7 210.95 211.45 Satt543 8 213.65214.47 Satt543 9 210.34 210.54 Satt543 10 216.56 217.05 Satt543 11178.15 178.78 Satt544 1 116.66 117.63 Satt544 2 147.56 149.12 Satt544 3154.30 155.84 Satt544 4 160.56 161.37 Satt544 5 163.56 164.34 Satt544 6166.54 167.72 Satt544 7 169.59 170.62 Satt544 8 132.15 132.45 Satt544 9151.41 151.61 Satt544 10 157.54 157.79 Satt544 11 172.44 173.53 Satt5451 242.25 242.85 Satt545 2 254.11 254.66 Satt545 3 260.29 260.86 Satt5454 275.32 276.23 Satt545 5 278.35 279.48 Satt545 6 281.59 282.30 Satt5457 284.46 285.32 Satt545 8 290.55 292.10 Satt545 9 293.64 294.56 Satt54510 266.36 266.78 Satt545 11 272.58 272.98 Satt545 12 287.84 288.04Satt545 13 296.79 297.29 Satt545 14 269.53 269.98 Satt546 1 332.22333.26 Satt546 2 341.33 342.06 Satt546 3 344.12 344.85 Satt546 4 355.29356.49 Satt546 5 358.46 358.66 Satt548 1 352.27 352.77 Satt548 2 359.73361.17 Satt548 3 362.57 364.12 Satt548 4 365.63 366.41 Satt548 5 348.57348.97 Satt548 6 351.34 351.54 Satt548 7 368.83 368.93 Satt549 1 225.08225.33 Satt549 2 233.92 234.80 Satt549 3 240.10 240.38 Satt549 4 243.05243.23 Satt549 5 245.94 246.80 Satt549 6 249.00 250.00 Satt549 7 255.38255.88 Satt550 1 223.78 224.73 Satt550 2 226.66 227.46 Satt550 3 241.76242.85 Satt550 4 244.72 245.71 Satt550 5 229.84 230.24 Satt550 6 220.91221.36 Satt550 7 232.80 233.35 Satt550 8 238.96 239.16 Satt551 1 230.50231.57 Satt551 2 236.85 237.63 Satt551 3 242.75 243.73 Satt552 1 160.66161.67 Satt552 2 175.66 176.68 Satt552 3 181.73 182.80 Satt552 4 184.98185.75 Satt552 5 169.80 170.62 Satt555 1 249.45 250.30 Satt555 2 271.56272.75 Satt555 3 274.87 275.78 Satt555 4 277.96 278.68 Satt556 1 135.95136.83 Satt556 2 148.62 149.94 Satt556 3 151.96 152.78 Satt556 4 184.68186.12 Satt556 5 197.30 197.99 Satt556 6 155.32 155.84 Satt556 7 164.34164.82 Satt556 8 168.22 168.69 Satt556 9 176.93 177.90 Satt556 10 188.45188.65 Satt557 1 201.89 202.90 Satt557 2 204.90 205.90 Satt557 3 211.20212.01 Satt557 4 214.17 214.97 Satt557 5 220.29 221.36 Satt557 6 208.42208.93 Satt557 7 193.45 193.75 Satt558 1 220.38 221.46 Satt558 2 229.54230.55 Satt558 3 235.38 236.66 Satt558 4 238.68 239.70 Satt558 6 233.05233.42 Satt558 7 241.86 242.45 Satt558 8 245.21 245.41 Satt560 1 231.06231.97 Satt560 2 237.11 238.91 Satt560 3 243.15 244.52 Satt560 4 246.20247.40 Satt560 5 276.80 277.67 Satt560 6 286.02 286.83 Satt560 7 273.70274.30 Satt560 8 271.25 271.56 Satt560 9 283.06 283.41 Satt560 10 250.10250.21 Satt560 11 253.13 253.24 Satt563 1 166.45 167.82 Satt563 2 169.65170.82 Satt563 3 236.85 237.83 Satt563 4 173.53 174.49 Satt563 5 181.49182.95 Satt563 6 212.36 213.47 Satt563 8 160.42 160.99 Satt563 9 164.34164.54 Satt563 10 206.30 207.41 Satt563 11 185.15 185.55 Satt563 12209.63 209.83 Satt565 1 299.47 300.33 Satt565 2 302.62 304.63 Satt565 3327.31 328.46 Satt565 4 351.34 351.84 Satt565 5 330.48 331.06 Satt566 1356.49 357.42 Satt566 2 359.44 360.23 Satt566 3 362.12 363.07 Satt566 4365.18 365.94 Satt566 5 374.11 375.06 Satt566 6 376.96 377.90 Satt566 7367.97 368.33 Satt566 8 379.97 380.17 Satt566 9 385.90 386.30 Satt567 197.45 97.95 Satt567 2 112.41 113.36 Satt567 3 115.25 116.20 Satt567 4118.56 119.10 Satt567 5 110.01 110.51 Satt567 6 98.05 98.15 Satt568 1345.38 346.08 Satt568 2 350.62 351.84 Satt568 3 357.02 357.22 Satt568 4354.12 354.42 Satt569 1 103.33 104.23 Satt569 2 115.60 116.30 Satt569 3121.47 122.55 Satt569 4 124.56 125.31 Satt569 5 127.58 128.38 Satt569 6130.63 131.38 Satt569 7 133.70 134.10 Satt569 8 106.59 106.99 Satt570 1108.50 109.05 Satt570 2 111.41 112.41 Satt570 3 114.48 115.25 Satt570 4126.57 127.58 Satt570 5 120.72 120.97 Satt570 6 130.01 130.21 Satt570 7132.94 133.17 Satt572 1 172.56 174.08 Satt572 2 175.86 176.53 Satt572 3178.87 179.50 Satt572 4 169.89 170.45 Satt573 1 167.36 167.56 Satt573 2163.84 164.82 Satt573 3 175.96 176.53 Satt576 1 261.38 261.88 Satt576 2288.75 290.25 Satt576 3 342.10 342.99 Satt576 4 350.48 351.84 Satt576 5361.83 363.07 Satt576 6 365.11 365.94 Satt576 7 353.64 354.12 Satt576 8356.69 356.89 Satt576 9 359.09 359.59 Satt577 1 312.82 313.96 Satt577 2316.01 317.25 Satt578 1 178.37 179.35 Satt578 2 181.29 182.25 Satt578 3199.89 200.39 Satt578 4 202.39 203.00 Satt578 5 199.49 199.79 Satt578 6184.78 184.98 Satt581 1 138.39 139.45 Satt581 2 148.06 149.22 Satt581 3151.21 152.43 Satt581 4 161.07 161.27 Satt582 1 333.83 334.34 Satt582 2336.72 337.31 Satt582 3 339.75 340.19 Satt582 4 318.76 319.16 Satt583 1122.06 123.05 Satt583 2 146.99 147.86 Satt583 3 150.17 151.21 Satt583 4180.82 181.64 Satt583 5 125.56 125.76 Satt583 6 131.58 131.95 Satt583 7137.89 138.09 Satt583 8 141.10 141.30 Satt583 9 144.15 144.60 Satt583 10113.46 113.66 Satt583 11 116.50 116.66 Satt583 12 153.80 154.00 Satt58313 156.84 157.04 Satt583 14 159.96 160.16 Satt583 15 168.99 169.19Satt583 16 198.89 199.19 Satt583 17 202.09 202.39 Satt583 18 222.89223.39 Satt584 1 171.59 172.56 Satt584 2 174.49 175.46 Satt584 3 180.77181.32 Satt584 4 195.43 196.42 Satt584 5 183.72 184.68 Satt584 6 186.91188.85 Satt584 7 198.89 199.89 Satt586 1 318.76 319.77 Satt586 2 337.12338.01 Satt586 3 342.99 343.79 Satt586 4 348.57 349.50 Satt586 5 354.14355.05 Satt586 6 331.18 331.77 Satt586 7 334.15 334.54 Satt586 8 340.05340.84 Satt586 9 351.34 351.94 Satt587 1 168.02 169.19 Satt587 2 170.97172.20 Satt587 3 173.93 175.05 Satt587 4 176.93 177.90 Satt587 5 129.10129.61 Satt587 6 155.84 156.34 Satt590 1 317.18 318.26 Satt590 2 323.30324.81 Satt590 3 332.86 333.55 Satt590 4 335.78 336.87 Satt590 6 339.25339.75 Satt590 7 266.88 268.52 Satt590 8 272.88 274.62 Satt590 9 264.22264.44 Satt590 12 314.28 315.21 Satt590 13 304.83 305.16 Satt590 14320.48 321.06 Satt590 15 300.73 301.25 Satt590 16 311.10 312.00 Satt59017 326.64 327.60 Satt590 18 329.87 330.88 Satt591 1 137.43 138.44Satt591 2 140.52 141.60 Satt591 3 150.27 151.59 Satt591 4 144.63 144.83Satt591 5 119.20 119.50 Satt592 1 236.85 237.43 Satt592 2 242.75 243.53Satt592 3 245.71 246.85 Satt592 4 254.76 255.87 Satt594 1 122.50 123.25Satt594 2 134.74 135.44 Satt594 3 141.10 141.60 Satt594 4 144.33 145.61Satt594 5 147.56 148.82 Satt594 6 166.22 167.36 Satt594 7 172.56 173.63Satt594 8 157.04 157.64 Satt594 9 160.06 160.46 Satt594 10 178.20 179.13Satt594 11 138.09 138.29 Satt594 12 169.19 169.59 Satt595 1 340.05341.13 Satt595 2 342.99 344.85 Satt595 4 345.98 346.38 Satt596 1 257.73258.83 Satt596 2 260.72 261.88 Satt596 3 266.68 267.70 Satt596 4 272.86273.90 Satt596 5 275.88 277.15 Satt596 6 281.80 282.91 Satt596 7 264.17264.64 Satt596 8 270.10 270.65 Satt596 9 279.43 279.58 Satt596 10 251.92252.12 Satt597 1 144.68 145.51 Satt597 2 147.86 148.82 Satt597 3 160.26161.37 Satt597 4 157.34 157.90 Satt598 1 163.36 164.37 Satt598 2 175.46176.43 Satt601 1 344.86 345.93 Satt601 2 347.84 348.57 Satt601 3 342.36342.66 Satt601 4 351.02 351.22 Satt601 5 356.39 356.59 Satt601 6 336.72336.92 Satt602 1 336.21 336.82 Satt602 2 339.14 341.14 Satt602 4 342.86343.92 Sct_010 1 107.29 108.10 Sct_010 2 99.45 99.95 Sct_010 3 105.44105.64 Sct_026 1 328.36 329.37 Sct_026 2 330.38 331.37 Sct_026 3 326.64326.84 Sct_026 4 333.80 334.25 Sct_026 5 317.50 317.65 Sct_028 1 236.07237.33 Sct_028 2 244.12 245.21 Sct_028 3 248.10 249.20 Sct_028 4 242.10243.23 Sct_034 1 126.67 127.58 Sct_034 2 128.83 129.61 Sct_034 3 130.63131.65 Sct_046 1 260.66 262.28 Sct_046 2 267.69 268.52 Sct_046 3 275.88276.65 Sct_046 4 277.89 278.68 Sct_046 5 284.15 285.73 Sct_046 6 286.23287.83 Sct_046 7 288.24 288.75 Sct_046 8 292.89 293.76 Sct_046 9 298.60299.83 Sct_046 10 290.05 290.75 Sct_046 11 270.03 270.55 Sct_046 12274.10 274.62 Sct_046 13 282.71 283.71 Sct_065 1 163.26 164.40 Sct_065 2165.27 166.40 Sct_065 3 171.12 172.18 Sct_137 1 269.32 270.03 Sct_137 2273.08 274.10 Sct_147 1 130.11 131.03 Sct_147 2 136.28 136.93 Sct_186 197.45 97.95 Sct_186 2 105.24 105.74 Sct_186 3 111.36 111.91 Sct_186 4113.36 113.80 Sct_186 5 115.35 115.70 Sct_186 6 117.36 117.63 Sct_187 1313.34 314.19 Sct_187 2 311.40 311.90 Sct_187 3 315.21 317.35 Sct_188 1242.25 243.23 Sct_188 2 252.23 253.24 Sct_188 3 254.76 254.96 Sctt008 1159.36 159.86 Sctt008 2 162.36 163.36 Sctt008 3 165.42 165.62 Sctt008 4147.56 147.76 Sctt009 1 213.93 214.97 Sctt009 2 219.71 220.86 Sctt012 1343.92 344.72 Sctt012 2 351.84 353.02 Sctt012 3 355.05 355.55

APPENDIX II SSR Markers: Table of Primers 9 pages Marker NameLeft Primer Sequence Right Primer Sequence Pigtail Repeat Sac1006CAATCAGGTTAGTGGTCCTACC CAAAAGGTTTTCAGTGGTGG GTTTCTT 2 Sac1677AAGTTCACAGCACCTGGACCAT CCAATCCCTACCCTGATTGCAC GTTTCTT 2 Sat_084GAAACAAACACCCCCAAGGTGA GGAAGTGGATTTGGAGTTGGGA GTTTCTT 2 Sat_090CCAATCGTGTCTTATCCTCGGG AACCCATAACCTGCTCGTCTCA GTTTCTT 2 Sat_104TTCCAGACAGAACCCAAGTAGCC AGGCGGGTTTGGAGCTGTTTAC GTTTCTT 2 Sat_110AACATTTTTCATCGCTTTTCTTAG TCTTCTCAGGAACTTGAATTACTCA GTTTCTT 2 Sat_117AAAGCATTTTTGGCAGTTTCTTGT GGAATGTCCCAAGTGTCAGCAA GTTTCTT 2 Satt020TGGACAGAATGGAGAAAGAAATGTG TGAAAATAAGTGGAAAGAAAAAGGAAAA GTTTCTT 3 Satt040CCAAGCCAAACAAAGAATCACA GTCCCCGTTCCTCAAACACCTT GTTTCTT 3 Satt042GCTTGCTATGATTGATTGATTGATTGA TGCACACTCACTTGGTCTTACACA GTTTCTT 3 Satt050AGAACGGTGTTAAGATAGAATAGTT CTGTTGGAAAATGATGCGTGGC GTTTCTT 3 Satt066GGGAAGCTTAATAATGAAAATGACAC TTGATCACTTCTGTAACATTC GTTICTT 3 Satt092AACCGATGCTTTTTTCGCCTTT CTTTGTTGTTTGGTCAAGGCCC GTTTCTT 3 Satt102GAAGAAGGCTCAGCAACACCTTG TGCCGAAATAAGTGAGAGCATAGAA GTTTCTT 3 Satt108CACCCAATCTTGCCTTTGAAACA TGTTAGGTATGGGATTTAGGGTTTTGA GTTTCTT 3 Satt109TAGACACTTTCAGGTTAAAATATAAC TCCACTCACTGTATGTCTTCCCTTG GTTTCTT 3 Satt111CAACTTGTCACTTACACATAGTTTAG TTGAGTGCTCTTGGTGTTTTCCT GTTTCTT 3 Satt115ATTTCGCTCGCAAACACAAGGT CCAAACACCCCAGTATAAAAAATGGA GTTTCTT 3 Satt122TCAAGAAATACAAGTGCAAGAAAGACC GGATTTTGAGTTGCTCCAAGGC GTTTCTT 3 Satt127AATTTTCGCTTGTGAACCCTGC TGTATGATCCATCCTCTGAAACCG GTTTCTT 3 Sat1129TTCAGTACAAGTCGGGTGAATAATAATA TCACATGTTCGGGACTTAAGGTAT GTTTCTT 3 Satt130TGGGACTCTCACACACGGAAAA TGAATGGCTAAAAACGTGATTTGGA GTTTCTT 3 Satt131TGGGAGTCAATTTCCCATTATCA GGACCTTCCCTTTCCCATGACT GTTTCTT 3 Satt133TGGATTTGAAACCACAAATAACAACAAC AGCGATGGTTGAAGAAAGGGTC GTTTCTT 3 Satt138AAAAGGGGGACATTTTTCCACG AGAGAACGGGCGATTTATGGCT GTTTCTT 3 Satt142CATTAGGGACAACAACAGCGTTT ATGTCGCCACTAGGCCAATCAG GTTTCTT 3 Satt144CGTCGCCATCACTATGAGAA CCATCTTGAGCAGAGTTTGAAGTT GTTTCTT 3 Satt146AAGGGATCCCTCAACTGACTG GTGGTGGTGGTGAAAACTATTAGAA GTTTCTT 3 Satt147CCATCCCTTCCTCCAAATAGAT CTTCCACACCCTAGTTTAGTGACAA GTTTCTT 3 Satt150TGACGAAGCTTGAGGTTATTCG TCAAGCTTATGCTCTATAGGCT GTTTCTT 3 Satt151GCCAAGAAGATAACAAGCTCGGC GACCAAAATTCAAGGCAGTGACAA GTTTCTT 3 Satt153GGGTTATATCAGTTTTTCTTTTTGTT CCATCCTCGTTAGCATCTAT GTTTCTT 3 satt155AGATCCAACACCTGGCCTAAT GCTGCACAATTCATTCCATTT GTTTCTT 3 Satt156AACAAAACTAGCCCATAGAAACATTGA CCCAGGGACTTACTTTTTTCAGTTT GTTTCTT 3 Satt159GAAATGCCCAGAAAAACCTAATAAC TGAAGCAACAAAATAGAGGAATAGAG GTTTCTT 3 Satt165GCCGTGAAGACAGTTGATCGTT CATTACTAGGCGTGTGTTGTTTCAA GTTTCTT 3 Satt166CAGTTGATTTTTGTTTTTCGGCA CACGCGCATCAGCTTTGTAGAG GTTTCTT 3 Satt168TTGTCCAAACTTGCAGGGAACA TCTTTTCACCATTCTCCAACCTCA GTTTCTT 3 Satt172TGGATGAGTTCTATTGGGGTAGTCG TCCTTTCTCCCATTTTTTTTGGG GTTTCTT 3 Satt175GACCTCGCTCTCTGTTTCTCAT GGTGACCACCCCTATTCCTTAT GTTTCTT 3 Satt176CGCAATCCCAATTCACCTCTTC AATTGTTAGGGCGCGAGAAACA GTTTCTT 3 Satt181GAACCCCGTTTCAACATTTTATGA CTAGCCAAGGGAGAGAGGAGCA IGTTTCTT 3 Satt183GGTCGTTAAGCCCACTTTGAGA CCTCACACCAACCAGCACAAAA GTTTCTT 3 Satt186TGCAGCTTTCACTAATCGTCAGAA CAATTTTATGATTTGCTTTTGAAGGGA GTTTCTT 3 Satt190GGGAGTGTGAACTTACATTGTCT GGGCCTTGAATTTTGTGCTAT GTTTCTT 3 Satt191GCGATCATGTCTCTGCCATCAG CCTCTTGAAACCGTGAAACCGT GTTTCTT 3 Satt193GAAGGGATATGGAGAGTGAGAAATTAGA TCTTTTTCTTCATTTTTGTTCGCA GTTTCTT 3 Satt195AAAAATTGTGTAAACGAAAGATGGGA AAACTGCTCTGAAGTGCGACACA GTTTCTT 3 Satt196TGAGCCCAACCTCCACATCTTT TGTGAATAAAAGAAATCCCCATTGA GTTTCTT 3 Satt197CACTGCTTTTTCCCCTCTCT AAGATACCCCCAACATTATTTGTAA GTTTCTT 3 Satt199GCGGTAAATGGTGAAAATCATTTATGGTT GCGTTTTCATACGGTGTTTTGCCTAT GTTTCTT 3Satt202 GGAATGCATGAGTATTAACCTCTTAT GGGCTAACGAACATGTAACTTATCAAC GTTTCTT 3Satt203 TGTCTTCCCAATCCATCTAATCTAATCA GCACTACAATATGTGCATGAATTTTTCTGTTTCTT 3 Satt204 CCAATTATGTTTCTATGCCATCTTGTT TCTCCTTACTCACTCCATTGGCAGTTTCTT 3 Satt209 TGTGGATAAAAGCCATCTCTAACAAA TCCATGTCACCAACCACAAAAAGTTTCTT 3 Satt212 TCGATACCAATCATCCAATCCAAA CGTATCCCTTTCCCATACGTGGGTTTCTT 3 Satt213 TCCTTAGACCTCTCGCGCAAAC ATGACGGGATGAAAGAGCCAAA GTTTCTT3 Satt216 TACCCTTAATCACCGGACAA AGGGAACTAACACATTTAATCATCA GTTTCTT 3Satt218 ACGTATGCGAAGGAGGGAGATG CGGAAAAGATATACCGAGTGAGAAAA GTTTCTT 3Satt219 TTGGCTCTTGTGTAAGTGGCCC TTCTAAGGATTTGTGGTAATCGGC GTTTCTT 3Satt220 TTCAGTCCCTTGGTGGTTCCAA CATGGAGAAAAGAAGAGGAGGGA GTTTCTT 3 Satt221CATGGTCCTTGGGTGCAATTTA AAGCAAACCATTATCTTCATTGGTG GTTTCTT 3 Satt225AAAAATGTGTTAGAGCTTGTGTTGTTA GCCACACTATTCCAGCCACTAC GTTTCTT 3 Satt227ATGGTGCAGTGTTGCAGGTTGT AGTAGTCCAAACCGAAACGCCA GTTTCTT 3 Satt228GACCACAACTTCTTTTTTGTGAATGG TCAATTCGGTCAAAAGGCTTGA GTTTCTT 3 Satt230CCGTCACCGTTAATAAAATAGCAT CTCCCCCAAATTTAACCTTAAAGA GTTTCTT 3 Satt233AGAAGCATACTCGTCGTAACACTATCC AATGGATGGACCTGTCAGTCTGC GTTTCTT 3 Satt234GAGCAGGACATTTTTTTTATCCTTGA TGCTTCCATTAGTCTCTCATCCTCC GTTTCTT 3 Satt236TGCTTGAGGTTGAAGGAAATGC TGGAAAAAGATAACGTGTGTTTGCAG GTTTCTT 3 Satt240TCCTTGGGGTGAATTGTTTTTCA CCTTCTTTTGACATGGGGCCTA GTTTCTT 3 Satt242GCGTTGATCAGGTCGATTTTTATTTGT GCGAGTGCCAACTAACTACTTTTATGA GTTTCTT 3Satt243 AATGCTTTGGTCGTTGCATTG TGGTATCGGGAGATTTTTTCAGC GTTTCTT 3 Satt247CAGCGTCTGCATGATAGCGTTT TTTTTCGTCAGCACATTATTACACATTT GTTTCTT 3 Satt249TGCAAATTGTTATTGTGAGACTGAATGA GGCCAGTGTTGAGGGATTTAGAA GTTTCTT 3 Satt250CGCCAGCTAGCTAGTCTCAT AATTTGCTCCAGTGTTTTAAGTTT GTTTCTT 3 Satt251CCTCCACCCCCTTCCCACCCAAAA GGTGATATCGCGCTAAAATTA GTTTCTT 3 Satt255AGCGTCGTCTGGCTAGGTCTGT GGAAACCCTGTCATTTTCGTGC GTTTCTT 3 Satt256GTCATTGGTCTCAAACAATCTTCAT AGAGTTCTCAGTCCGCCAGCTC GTTTCTT 3 Satt257GCGACTTTCTTTTCAATTTCACTCC GCGCAATTGTCACCAACACAT GTTTCTT 3 Satt258CACTTTTTCACTGTCTCCCCCC CCCAAACCAACAAGCAACAACA GTTTCTT 3 Satt259TGGGCCATTTGGGCAGCTCGACT ATTCACACGCATCTGGAATAATA GTTTCTT 3 Satt262GCGCCCCATTAATGTTAACACA GCGGAGTTCAACGCATTCACCTT GTTTCTT 3 Satt263GGAGAGAATCCATATATATTGAATTGC TGAAAGCAAACGAGACTCATGGA GTTTCTT 3 Satt264CCTTTTGACAATTATGGCATATA GCATAGAAGGGCATCATTCAGAT GTTTCTT 3 Satt265CCTGTCAAATTGGCTGATGCAA GCTAGATGAGGAGACCATTGCACTT GTTTCTT 3 Satt266TTTTACCAAACAAATTAAACTGCGTCT CAAGAGGTTGTTGTAAGAGTGATCTCG GTTTCTT 3Satt267 CCGGTCTGACCTATTCTCAT CACGGCGTATTTTTATTTTG GTTTCTT a Satt270GGTGTTTCAAGTTTCAACACCAT CAGTGCATGGTTTTCTCATGTACC GTTTCTT 3 Satt272ATGACAAGGAAAAATCAATCAAC GCTGCTGTTAAGAGTGTTTG GTTTCTT 3 Satt274GCGGGGTCAATTAGTTTTCGTCAGTT GCGCACGGTATATAATCGAACCTAT GTTTCTT 3 Satt279GCGCAAAAGGACGCCCACCAATAG GCGGTGATCGGATGTTATAGTTTCAG GTTTCTT 3 Satt280TCTGCTTATTCATTGTGTGCGTG TGTCTCCATGCTGTAACACGTCAA GTTTCTT 3 Satt282TGGTATATGTTTTTGCGGGACAA CGCCAAAGATGCAAGACAGTTG GTTTCTT 3 Satt284AGGTGGGCTAGGAGTGACCACA TGTAATTGCTGTTTTGGTTTCATTTC GTTTCTT 3 Satt285GCGACATATTGCATTAAAAACATACTT GCGGACTAATTCTATTTTACACCAACAAC GTTTCTT 3Satt287 GGGGTGAATGAATGTCAAGATGA GCGCGAGGTATCAACACAATTACT GTTTCTT 3Satt292 GCGGAATTAGAACTCCAGTAAAGA GCGAGGCCAACATTGAAAAGT GTTTCTT 3 Satt295TTAGTGGATCTACACTAAACTAATCCG CGTACCATTGGACAACTCTGTAATTCAA GTTTCTT 3Satt299 AGGCATTTCTGGCAAGGGTATG TGGTGAATCAATCTCCTCTAAGTGC GTTTCTT 3Satt300 GCGCCCACACAACCTTTAATCTT GCGGCGACTGTTAACGTGTC GTTTCTT 3 Satt303ATGGGCTATGGGAGGAGGTGTT CCACACGGGACTTTCCATTTTC GTTTCTT 3 satt304CCAGTGCAGTTTTACATGAACTT TATATGTAATGACCCCCATCATG GTTTCTT 3 Satt307GCTGGCCTTTAGAACGTCTGACT CGTTGGATTCGACTTTTTGGGA GTTTCTT 3 Satt311CCACAAAAAGATGAAACAAAATAG TTGAAGCTCAGGCTGTGATGAAT GTTTCTT 3 Satt314GCGGAGATTGGAACCTACTCATTC GCGGGGACCAAAAATTCAAAA GTTTCTT 3 Satt319AACAATTTGATGGTCGGGGTTG TAGGGGAACCGATTTGGTGAAA GTTTCTT 3 Satt321CACCGTCGTAAAAACTGTGTCGT GCGTGTCAAAGAGTTTTAGACATC GTTTCTT 3 Satt322GAAGTGCTAGTTCTCACGGTGTGG TTGGAGGGAGGTTTTGGAAGGT GTTTCTT 3 Satt326TTCGCTGCATAATTTTTAGCATCA CCAATCTTTTTGTTAGTTCACCTTCCA GTTTCTT 3 Satt327AAAGAGACACCCAAAAGATAACAAACA TTCGTAGCAATGTCACCACCTTGT GTTTCTT 3 Satt328TGACCACCATGAGTTGATT GGGGGTGGCTTTTAGATTC GTTTCTT 3 Satt329GCGGGACGCAAAATTGGATTTAGT GCGCCGAATAAAACGTGAGAACTG GTTTCTT 3 Satt330GTGACCCTCCATTCCACAACAA TCCTTGCCTTTTAGTTGTTCGGT GTTTCTT 3 Satt331GCAGAGTCCCCCCTAAATATAG CGGGAACAACCACACTCTCCATT GTTTCTT 3 Satt332GTGAATCATCCAGGGCTTGC TCCTTCACTTTCAAAAACAAAAACAA GTTTCTT 3 Satt333GCGAATGGTTTTTGCTGGAAAGTA GCGCAACGACATTTTCACGAAGTT GTTTCTT 3 Satt334GCGTTAAGAATGCATTTATGTTTAGTC GCGAGTTTTTGGTTGGATTGAGTTG GTTTCTT 3 Satt335CAAGCTCAAGCCTCACACAT TGACCAGAGTCCAAAGTTCATC GTTTCTT 3 Satt336AATTGGAGTGGGTCACAC TTCCCGGAAAGAAAGAAA GTTTCTT 3 Satt338GCGCCCAAGTATTATGAGATATTTGAT GCGATAATTTTAAAACTGGACCA GTTTCTT 3 Satt339CTTTGTTTGGTTGGTGATAAGTTTCTA AAGCAGTTCCTCTCATCACGTAACA GTTTCTT 3 Satt343AATTGTTAGGGCGCGAGAAACA CGCAATCCCAATTCACCTCTTC GTTTCTT 3 Satt346GTTCGGAGGGAGGAAAGTGTTG CCATAAAACATAGCAACTGTCGTCTC GTTTCTT 3 Satt347TGTTCAATCGACAAATAAGGGTGC TCCCAGGGTAAAAGTTCAAGTTCA GTTTCTT 3 Satt348CTTTACTTAGTAATGGTTCCCACAG TCGATGTTTCTCCCTCCCTTAGA GTTTCTT 3 Satt352CCAGATTTTACGTTAATGTTTTAGTTTTA TTGCACATGGTCCTTGGTTGAT GTTTCTT 3 Satt353CATACACGCATTGCCTTTCCTGAA GCGAATGGGAATGCCTTCTTATTCTA GTTTCTT 3 Satt355GCGTCCCAGGACATCATCATCATC GCGTAGCGTGTTATTTTGTGTTTG GTTTCTT 3 Satt356TGCTGCTTGTGTTTGGTTGATCT CATATCCTGCCCCCCCAATTAT GTTTCTT 3 Satt357AATCCCCAAACAAACGCACAGT TGACATCTAAGTCCAGAAATCAAAGCA GTTTCTT 3 Satt358GCGGCGCTTTATGTAACAATACGATTT GCGAGTAAAAGCAGAGTGCGGAGTA GTTTCTT 3 Satt359GCGAGAAAATAATCCTGCTCAAG GCGTTTAAGTCCAATAACAAAGATAAC GTTTCTT 3 Satt361TCGGGAGACACTAAAGGCACTG TCGTTGACACACAAAAAAAGCGA GTTTCTT 3 Satt364GCGGCATAAGTTTTCATCCCATC ATCGGGTCATGACTTTTGAAGA GTTTCTT 3 Satt367GCGGATATGCCACTTCTCTCGTGAC GCGGAATAGTTGCCAAACAATAATC GTTTCTT 3 Satt369GGAGAAAAACATCCAAAGAAATGTG CAAGTGGATTGACACACTAAGGTTTGA GTTTCTT 3 Satt372GGATAATTTTTTCATCATCACAATTTATG CACAAAAGACAGGAGATGTGAGCAA GTTTCTT 3Satt373 TGGAAATACTGAATGAAAGCATATTGG TTGTAGACGACTTGTGGTTCGATTC GTTTCTT 3Satt378 CCATTGGGATCGAGAATAATTGATG AATGAAGAAAATGTGAATTTGAAACCA GTTTCTT 1Satt380 AGAGTAATGGCTCCTGCTCCGA TTCCCTTTTTCTAACTCTCCTTTTTCA GTTTCTT 3Satt383 CGATCTAACACGCATATTTCCTCTGA TGTCTTGGTGCAATACCTGACATTT GTTTCTT 3Satt384 TGGGGGTCAATTTTAATTTGTGC ATTTCCCTTTGACCCACCTCTGTTT GTTTCTT 3Satt385 GATTCTCTTCATTCTAATACTCGTTT CAACGATCCCAGCTCACAGTTT GTTTCTT 3Satt387 GCGTTACGTTTCACTATTTATTTAACAT GCGGCAGGCTAGCTACATCAAGAG GTTTCTT 3Satt389 GCTGGTGTATGGTGAAATCAAATTACT TTTCAAACAAGGAAGAAACCTCTTTTT GTTTCTT3 Satt390 TGATATTGTTTTGTGTGAAAGATGCAC AAGTAACACTGTGGCGGCATCC GTTTCTT 3Satt391 TGCTCAAAGGGTCAATTTCTTTCC TGTGTAATTTCTATCACCTTATTGTGCC GTTTCTT 3Satt393 CCAAGCCCATAAACGAAATAAAACA TCCTTTGGCTCGGCCTATGTAA GTTTCTT 3Satt398 CAGTGCTCATATCAAATTAAAGTGG CGCGGACTCAGTTAAACCGTAT GTTTCTT 3Satt399 TTTCAACCACCAAGCCAACCTT CGTTCAATAGTTCCTATGATGGACGA GTTTCTT 3Satt406 TGCAGCATGTGTTTTAGGCTTTC GCATTGCACGTCGATTTTAGGG GTTTCTT 3 Satt409CCTTAGACCATGAATGTCTCGAAGATA CTTAAGGACACGTGGAAGATGACTAC GTTTCTT 3 Satt411TGGCCATGTCAAACCATAACAACA GCGTTGAAGCCGCCTACAAATATAAT GTTTCTT 3 Satt412ACTGGCGCTGACCTTAAATTGC TCCTTTTAATTCTAACATTGAGACAGCA GTTTCTT 3 Satt413TGTTTTTAAGTAATCCGGTGAAATAGCA TCTGTCCCAAAAAAGAAAGAAGATATG GTTTCTT 3Satt414 TCACATCACAAGTTTCATAAATGCTG CAATCTTTAATGCTCTGGAGTTTGAGA GTTTCTT 3Satt415 GCGTCTCCCTTAATCTTCAAGC GCGTGTGACGGTTCAAAATGATAGTT GTTTCTT 3Satt416 AGATTAAGAGAAGACGAGAGTTTTA GGGTAATTTATCTGTGTGATTGTTC GTTTCTT 3Satt417 GCCAGGTGCTCACTTCTTGCTA TTGCTTGGGATTTTCATTTTATTATAGG GTTTCTT nSatt418 CAGGAGAAAAAAGGAAAAGAAAAGCA GGTCAAAGAATAAGGGATTTGCCTC GTTTCTT 3Satt420 AGGTTTTGCTTCTTGAAAGTGAAGAG TGGGACTTTGATTTTTGGAATACCC GTTTCTT 3Satt421 GTCTCCGTTCAAAGCTTCTTCTTC CCAGAGAAATTATTGGAGTGGCAA GTTTCTT 3Satt422 GAGGGGAGGTAAAAAGTCGGGA GCGCAAAGAAATTCCCATCCTA GTTTCTT 3 Satt423CGCTTGGGTTCAGTTACTTGGTG GGGAGAGGTTCAGTTGGGGAAT GTTTCTT 3 Satt426GCGCCCATTATTATTTTCCTTGAATTGT GCGAATGCCTAATGTGATGATAAAAAAATA GTTTCTT 3Satt429 GCGACCATCATCTAATCACAATCTACTA TCCCCATCATTTATCGAAAATAATAATTGTTTCTT 3 Satt430 AATGCAAGAAGCAATCAGCAAGA AATAATGGAGTGACCCGCTGCT GTTTCTT3 Satt431 TGGCACCCTTGATAAATAAGAGAAGG TGGTACGAGTGGCACGAAATGT GTTTCTT 3Satt432 AATTGAACCACTCAGCCAAGCC GCCATCCTTTCCTTTCAACCAA GTTTCTT 3 Satt433GTCTAAGATTTTAATTAGGGTGATTC TGTAGGCTATTGGAAGGGTGCG GTTTCTT 3 Satt436GCGTATAAAGAAAAACGAGCATATCAT GCGCTTATAAAGGCTTGTGAAAGACACT GTTTCTT 3Satt440 CAAATTGAAAAACACAAGAAAACACAA TGGTTAGTGCAATCTTGGCGGT GTTTCTT 3Satt441 AGCAACTAACCTTGGGCTTCAGTAA TGCACCCATCAATCACATTTTTG GTTTCTT 3Satt442 CCTGGACTTGTTTGCTCATCAA GCGGTTCAAGGCTTCAAGTAGTCAC GTTTCTT 3Satt444 AATTCCTTTGGCATTTTTGTTGC TTTTTTCTCACACCCATGCCGT GTTTCTT 3 Satt445TTTGACTTTTGAATCATTGCATCG CCGGTTGGCGTAATGTGTCTTT GTTTCTT 3 Satt448CACCACTCGTATCCTTCACAAGAGC GCCAGCAGCCTGTTCAGTTTTT GTTTCTT 3 Satt451GCGCAATTAAAAGGATAACTTATATC CCCCTCTTTGGCCCTCACACCTTCTC GTTTCTT 3 Satt452AAAATTCATGTCGCTGCGTTCA ATTTGAAGCTCTTGGTATCTTGGC GTTTCTT 3 Satt454TGAAAACCATGTCAAAGTAATGGCA CAACCATGATAAATGTGAGTGAGCTTG GTTTCTT 3 Satt455GGAAAGTTTTGTTACATGCCGGA GTCACAATTTCACGATCCCAAA GTTTCTT 3 Satt457TTTTAACTGGAGAAACCTGAGGGA TCCATTTTCCCTTTAGTCCAACG GTTTCTT 3 Satt460GCGCGATGGGCTGTTGGTTTTTAT GCGCATACGATTTGGCATTTTTCTATTG GTTTCTT 3 Satt461TTGCTTGCTGCACATGACTGAG AATTTCTTACGTTTCCATAGATTTCTCG GTTTCTT 3 Satt464TTGAAATAGTGGTGGGGTTGGG TGCCCTCTTCCCAAACTAGGGT GTTTCTT 3 Satt466TGTGTTGTGTGGTGTGGTGGTC CAACCACTGATTCAAGCCAACAA GTTTCTT 3 Satt467CAAAGTCCCCTTTCACACCTTTT CAATTTAAGCACGGTCATATTTTCTCA GTTTCTT 3 Satt469AAAGGGAAAGGAAGAATAAACCGA CGTGATGCAGTGAATTTTTTTCG GTTTCTT 3 Satt470TGCTTTTTCTCTTTGGCAACCC GGGTTACTTTTCTTTATCCTCCTCCA GTTTCTT 3 Satt471GCGCCCAAAACTATCTAGTAATTCTT GGGCTATCAAATTGACTAAAGCCAAA GTTTCTT 3 Satt473CCAACAACCAAATCAATCACTGC ACACTTGAATCATCGAGAGTTGCTAA GTTTCTT 3 Satt475GCTCCGGTCCTTCAACTGACCT TGTGCTGCTTGCTTCAATTTGG GTTTCTT 3 Satt476ATGTGGGTATGTTGCAGGCAGA TGGCCTGTGTTATATTACCGAACCAA GTTTCTT 3 Satt477GTTGGGAAAAGGTTACTACCATATC GGTCCGTATGCAATTCTTGACTAATA GTTTCTT 3 Satt478CAGCCAAGCAAAAGATAAATAATA TCCCCCACAAGAGAACAAGAAGGT GTTTCTT 3 Satt479GCGCTTTCAAAAAGTAACAATTAATGAAA GCGGGAATTGGTTAATCTCATCGTGAC GTTTCTT 3Satt480 GACCTTTCATCATCGTCCCCAC TGCGAAAAAGCAGAGTGACCAA GTTTCTT 3 Satt487TCAATTCATCACGGACCAGTTCA TGAGCATATTTTGATCCGATGCC GTTTCTT 3 Satt488GTGAGTTTCGGTGCTGTATTCC GTTGCTTTGTTATGTAATGGAAGTC GTTTCTT 3 Satt491CCTAAATTGATGAAAGGATACAAG GCCCCACAAATATTCAGAAGGTAA GTTTCTT 3 Satt492GTATCGTTCGCGTCTTGAGTC GCAGCGGTGTAGTTCGTTCTTTCT GTTTCTT 3 Satt493TTAACGGGAAAAAATTAAACCTACGA AAATCGTCTTTTGTGGCTGCCT GTTTCTT 3 Satt495TGGAGATTTAATATAGATGCCGCGA GCACCATGTTCTTTTTCCATCAAA GTTTCTT 3 Satt497GCGGTTTTGGATTGACTTTGTTGA GGCTCAATTAGAGCATGCAACATC GTTTCTT 3 Satt503CCGTGACTTTTGTTATCCTGAGTTCC CATGTTAAACGTCCACCCACCA GTTTCTT 3 Satt506GCGAATTGGCATACATAGTACC GCGTGAATTCGCCTAAGTTTAT GTTTCTT 3 Satt507TGCACCACTAATGTCCTCAGCC TCCCTACTCTCGTGTCGTTAGTTATTTT GTTTCTT 3 Satt508GCAATGGGTATTGATCGTGTCA AGTTACATTATTTTTGTCTTTCTGCCGT GTTTCTT 3 Satt509GCGCTACCGTGTGGTGGTGTGCTACCT GCGCAAGTGGCCAGCTCATCTATT GTTTCTT 3 Satt510GCGAGTTTCGCCGTTACCACCTCAGCTT CCCTCTTATTTCACCCTAAGACCTACAA GTTTCTT 3Satt511 TGCGACTTTACTGAAAACCTGGAA GGAAATGCTTCAAACCAACAAACA GTTTCTT 3Satt512 AACGTCTTCAAGTCAAGTGCCTACA GCCCACATAGTTTTCATTTTTCTCCA GTTTCTT 3Satt513 GCGCATCACAAGTTTTATAGATGCTGA GAGGTCTAGTGCTTTGGTAAGGTT GTTTCTT 3Satt514 GGGTACATTTTATTAAAAGTAAACACACC TGTCACACAACCAGTGTCTCAAAATC GTTTCTT3 Satt515 TGAACCTTGTCTGTTGATTTTTTTATGT CACACCCCAGGACCCATTAAGA GTTTCTT 3Satt517 TCTCCTACTTCTCTTTCTCCCGTTCA AAAGCGCACACAATGCAAATACA GTTTCTT 3Satt519 CCTGATTATATGTCTAGACAAACAT CAAGGTTACGAACTGCTCGAATAAG GTTTCTT 3Satt522 GAGATCACATCAAAGTCAAAACTGCC TGAGGAGGCAAGATGATCCAAA GTTTCTT 3Satt523 GCGATTTCTTCCTTGAAGAATTTTCTG GCGCTTTTTCGGCTGTTATTTTTAACT GTTTCTT3 Satt524 GCGAATTATCCAAAGATACACTTAGTC GCGGGTCTTACGAACGTGTCACATTATGTTTCTT 3 Satt526 ATATCGAAAATCGCGCATCTGG CGAACCCAAACCACAAAGCATA GTTTCTT3 Satt529 GCGCATTAAGGCATAAAAAAGGATA GCACAATGACAATCACATACA GTTTCTT 3Satt532 GCGCCAATATTATCATGCTTTATGT GCGTGTAAAAATCTTTGAATCTTGA GTTTCTT 3Satt533 AGTGGTCGTCACAACACTATCATAT CACCCATTATTGAAAATACAAGGACCA GTTTCTT 3Satt534 CTCCTCCTCCGCAACAACAATA GGGGGATCTAGGCCATGAC GTTTCTT 3 Satt536GCGTGGAATGAGACGTAACCAA GCATAATGGTCTAATAAAAGTGGAGACC GTTTCTT 3 Satt537CAAAACTTATGTGCAACACGACTTCA TGCTTTTGGAGGAACTTTGTCTCA GTTTCTT 3 Satt540AATGTAGCAATTTGACTGGCGAA CATTCAACCGTGATTGCGAAGA GTTTCTT 3 Satt543GCGGATCTAAGGATAATTCATTAA GGGAGCGGATCATTCGGTGAAA GTTTCTT 3 Satt544GCTATGGGAAAAGGATGTGTG GAGCTACCCGAGATGATACTC GTTTCTT 3 Satt545ATGTGATGGCATGTGAAATGGT GGATCAAATTGGGAAACACAAAGG GTTTCTT 3 Satt546CAGGGTATAGTTCAATTCAGTGAGCG CTCACATACATGGCAGCCGTAA GTTTCTT 3 Satt548CCTCTTTGTTGGTGGTTAAGTCTCC TGCCTTTAGCTGGTGGGAAAAA GTTTCTT 3 Satt549GCGGCAAAACTTTGGAGTATTGCAA GCGCGCAACAATCACTAGTACG GTTTCTT 3 Satt550TCGTCAATTAAGCAAAAATGTGAGA TTAGAGGTTTTCGGATGAGCGTG GTTTCTT 3 Satt551GAATATCACGCGAGAATTTTAC TATATGCGAACCCTCTTACAAT GTTTCTT 3 Satt552ACAAAAAGAAATCGAACCGGCA GTTTTGGTTGATCCGCATTGGT GTTTCTT 3 Satt555TGGCTTTGATGATGTTTGAGACAA TTTCATTACCGCATGTTCTTGGA GTTTCTT 3 Satt556CCCAGATACAGACAATAAAACCCGA TTATGTTCGTTCATCTCTGAAGCCT GTTTCTT 3 Satt557TCCACCATGTAATATGTGAAGTGGAT TTCTGTCCATTCTAGCTCACTAACCC GTTTCTT 3 Satt558CTCACACCCTTTCATTATCTA AAATCGCGCATCTAAATTTAC GTTTCTT 3 Satt560ACTCTATTTTATTATCGTGCAAGAA ACAATAACTTGTTTTGCACACTATT GTTTCTT 3 Satt563GATGACAACGTAGGCTAAAAA GCGCCCACATGATTTTGTACTGAT GTTTCTT 3 Satt565GATTCTATATCCATCGTGTTGCT TATGGTAAATATTAACCATTGTCCT GTTTCTT 3 Satt566CCCACTGTATCCTTAGTGTGCCA CCTCGTTTTATTCCGAAAGCCG GTTTCTT 3 Satt567GGCTAACCCGCTCTATGT GGGCCATGCACCTGCTACT GTTTCTT 3 Satt568CATTAACTAATAAGTTGTTGGTAGC TTAGATTCGGACACCGGTCTACT GTTTCTT 3 Satt569ACCAAATTGCTTCACGCATCC TCTTAATTTTTTTAGGAATGGCATCAA GTTTCTT 3 Satt570TGCTCATGTGGTCCTACCCAGA CGCTATCCCTTTGTATTTTCTTTTGC GTTTCTT 3 Satt572GCGGAGCATGTAAATCCAGCCTATTGA GCGGGCTAACTTATGTTACTAAACAAT GTTTCTT 3Satt573 GCGGATTTCGATTTGAATATACTTAC CCTGTGGCTGTTATACTATGCATATA GTTTCTT 3Satt576 TGGACACACACAAACACCTACAGAA GGGTGGCGTTGACAATGTTTTA GTTTCTT 3Satt577 AGCAAGTCTTGAGTCTTTTGTCT TTATTATCTAAACTTATATGTGCAT GTTTCTT 3Satt578 TCCCACGTCATATCCACTGCTC GCCTCCTAAGTCCGTACACAGCAT GTTTCTT 3Satt581 CCAAAGCTGAGCAGCTGATAACT CCCTCACTCCTAGATTATTTGTTGT GTTTCTT 3Satt582 CCGGTGATACTCCATACCAATAACA GGATTTGGTTTCTGTGTGCTGTG GTTTCTT 3Satt583 CCGAGCTAACAAAGGCGACCAAAT GGGGCACAAGCCACACTT GTTTCTT 3 Satt584GCGCCCAAACCTATTAAGGTATGAACA GCGGGTCAGAAGATGCTACCAAACTCT GTTTCTT aSatt586 ATGGCCGTCTCAAAAGAACTGG TGGGCACTTGCAGTCCAAATAG GTTTCTT 3 Satt587GCGAATGGTTGCTCAAATAATC GCGCAAACCGCACAAGTTTATGT GTTTCTT 3 Satt590GCGCGCATTTTTTAAGTTAATGTTCT GCGCGAGTTAGCGAATTATTTGTC GTTTCTT 3 Satt591GGCAGACTCGTAGAGCAATTTA TGTTGAAATTGACCAAAATTCCCA GTTTCTT 3 Satt592GCGAAGATTGGTCTTTTATGTCAAATG GCGGAGGAATACAAGTCTCTATTCAA GTTTCTT 3 Satt594GCGGTAACTCCTCGAGTOCCTCTCAAT GCGCCGCTAACAGACATCCAATA GTTTCTT 3 Satt595TGGTGATGGGAAGCAAACAAGA TGGATTTCACCCAAGAAAAAAGC GTTTCTT 3 Satt596CCATCCCTTCGTCCACCAAATA TCGACTACCCGTCGATTCCGTA GTTTCTT 3 Satt597TGCTGCAGCGTGTCTGTAGTATAATTT GGCACAACCATCACCACCTTATT GTTTCTT 3 Satt598CGATTTGAATATACTTACCGTCTATA CACAATACCTGTGGCTGTTATACTAT GTTTCTT 3 Satt601GTAACATTGGTTGTCATCTTTGTCTA ATCGAACTGTGACCGTCCCTTC GTTTCTT 3 Satt602GGAGGTTATCTAGTGGTATAGATGGT AAGGGAACAGAGTCGTGCTTCTTT GTTTCTT 3 Sct_010CCAAAAGCATTGAGAGTGGGGA CCAGCAAACCCCCAGGTAAAG GTTTCTT 2 Sct_026GAAACCCGAAACGCAAAATCTC GAAGAAAACGCGAATAACCCCA GTTTCTT 2 Sct_028CTCTCGCCGGTACAAAACACCT GCACGCAGACTCAAGTTCATTCA GTTTCTT 2 Sct_034TCACTCTCACAACTTCAATCTCTTTCTC AATAGTTGGGTCGTCGAAGGGG GTTTCTT 2 Sct_046CACGACTCTTCCTCTTCCTCCG TCCAACTTAACACAAGATCAGCGAA GTTTCTT 2 Sct_065CCCTGTGTTTCCCTCT GAAAAGTTTTATGTTCTGAGTG GTTTCTT 2 Sct_137GTGTTGCTCTTGGGAATCTGGC CCACACACACTGACACAGTAAAGCA GTTTCTT 2 Sct_147TCTCGACTCACGACTCA CCAAGGTCTCTCAGAGG GTTTCTT 2 Sct_186AAAATGAAAACACACAGAGAGAGAGAGA ACGGAGGCACTTCCCATTGTTA GTTTCTT 2 Sct_187AGCGAAATGTGTGGTCCAAGGT CAAAACGACATGACAAGGAAACTTCA GTTTCTT 2 Sct_188CGTCGAGAAAGGAAGAGAGGCA TCCTCCATAAAAATTAAAAACATTGGAA GTTTCTT 2 Sctt008GCGGAAACCATTCTGACGGATA GCCCCCAGACACAACATAATCA GTTTCTT 3 Sctt009GGAGGAACTTGGAAGGCATATCA CGAGAGAAGCAGAAGCAGAGGC GTTTCTT 3 Sctt012ACGGACAACGCTGGCACTAAG GAGAAAGGTGACGATGGACGCT GTTTCTT 3

APPENDIX III ASH marker primer sequences 2 pages Primer 1Primer 1 sequence Primer 2 Primer 2 sequence Marker Name Primer 3Primer 3 sequence P10355B-1 19392 CATGGTTTCTCTTATCTTAT(AG)ACATTGTTGCCAAG 19393 CAATTCATGGTTTCTCTIAT(AG)ACATTGT TGCCAAG 25400CACTGTCCCTGCTCCTGTTTCAAGTATC P10598A-1 14507CAATTCTTGTGGGTTGAAGCCTTGTTCTGAC 14505 GGAATCAACTTCTTCGTGAGTGGGTTGTTCP10615A-1 1125 TGGTGGCTATGGAAATCTCATGTGTGGA 4186CTCTCATTTACCAAACTCCAACATTTGATCA CC P10618A-1 14492CACACTGGTAGATGGGAAGCAAGAATAGG 14490 GAAGAAGATTCCACCCAGATCATCATCAG TAGP10620A-1 12099 GCTTGTGCAGCTCCAATCGGTGTAAC 12101GTGCAATCCAAGACATCTGGTTCGGAC P10621B-2 12158CAGCTAAACCTTACAAGGATGATTGGTCAAG 12159 CCCTGGACTGAAGTTGCCATAATGTATCP10623A-1 12153 GCTGGTTGGGAGAAAGCACTTCC 12154GAATCTAACATTACGCTCTGCTGGACTATC P10624A-1 14328GAGGGACTATGTGAAATGGAGAGGAGTG 14330 GTATGCTAAAAGAGGAGACTTGACTGGTGAGP10632A-1 12105 GATGAAGGAACCAACACTTGCATAACA ATTTG 12106TGTCAGCACTCCTCACTCATTTGCCGA P10633A-1 4187 CACTTAACAGGAGTGCTCCTGATCACCAG1183 GAGAACAAGGACAAATCAATAGGTGAGACGA AGAAA P10634A-1 12600CTCATCTGCTCAGAACCTTCAGTCAGTC 12601 CGGATCATGTCTAGTACATTAGAGATGCTTG TGP10635A-1 12779 CTACACTTCTAATGCCTATTTAGGTGTGCT TG 12780GTCATATCTAGGGAGATTTCTAACCAGTTG TC P10636A-1 15252TAGGCAGCGTGACAAACTGAGCATAGG 15258 GGGTTGATGTCCGATGGGTAAATGAAGTTGP10637A-1 14136 GCACTAATACTTTAGTTGACTTTTGAGGTGG TTGAG 14138GCTATGTGGTAGAAGTATATGAAAAGGTAGA TGACAG P10638B-2 15290GAATATACTAGCTTGATGCCTATTTGTTTCT AAACCC 15040AGCAGTCATACAATGCTCTTTATTGTGGTGA AG P10639A-1 15345GCTCATAGCCTGCTTCTTAATCTTGTTATTC TG 15347 CTATTTGTTTCAGGAGTTTCACAACCATCTCAAGT P10640A-1 15280 CATCTTGACCAGTCACCAATCTGAGTACAG 15281GGCTTCAGTGAGAAAGGTGGATCAAATGGA P10641A-1 15349CAACGTGTAAAATCAAGAGATTGAGCTTCT GG 15350 TCGGTGTGTTCAACTATGACTTTGGTTCTGP10646A-1 5642 CCCCCAACAAAACTAAAAATAGAACCCTCAA CAACC 6803GGTCCAAGAACATTATGATCTTGAACAATCT TCAC P10648A-1 14436TGGCTAGAGCATCAACACATCTATACCTTC 14333 CATTGGGCATGATTCTTGAATAGCCTTTTTA CCP10649C-3 12171 GAGGGCTATGTTTTCTTCTCCAGATGTGAG 12173AAGGTCGGCTTGGTGGTTAAAGGCAG P10651A-1 10414GCCTTTTATGCACATTTTTCCTGGGCATCTA AC 10415 CTCAATGTCATGGGATCAATTTGGAAATTCAATGACC P10782A-1 15294 GATTACATTAATTACCGCTATGACTATATC TTGGGAC 15296GCTACCTTCTCCATTGCTTCTATGTATTGG TC P10783A-1 15771GGGCATCACATACATGAAAACAACTACACT TG 15772 CAACAGCTCTCTTCCACCACAATCCTGP10792A-1 15805 GAGATTGGAAATTGTAGCTCTCTTTACTTGC TG 15807CTTTGAGGACTTATTTGGTTGTTATAGGCAT TTGG P10793A-1 15621GTGTTTCCCTCCATTTTTGCCAAAAGACAG 15814 TATACACACTAAGAATTCGCTCGCTGTACAAP11070A-1 16188 GGTCTAGACTTTCACTCAGACAAGGAAC 17490CAAAATACTACAGACCTAATTTGTAACTAAT TGCTCCC P11347A-1 9642GGGAAGAAGAAGAACACTCGGTACAGTAG 7693 AAGCTAGGAAATCCACACTCAAATTATCGACTTGTGT P12105A-1 23538 GACCTGAAGCAAAACGCCACCATTTCC 29311CGTTGGAGACGACGCCGTTTGATTAC P12198A-1 39113 CAGTCGACACGTCTTCTACTCC 39114TGGAAGGCATGTCGGAACTTG P12390B-1 21397 CCTTTTTGCCCTCACTTCATGCCTTCTATG23525 GCATAACCCAAGAGCTGGACTGACAAG P12391A-1 21215CAAGCTCTGCTGCCAGGTTAAGTGTTTC 21216 GACTAGAACAAATTGGGGCTAGTGTGTTTG AGP12392A-1 21494 TGGACTTGCGGGACTATGCCTTAGAG 21495GCAGGAACACGTTCGTAACCATCAACTG P12394A-1 22161GTCCCTTTCTGAACCACTTAAAGAGTCAAC AG 22163 GGCATAGTGAGTTGAATACCAGGAGGAATCP12396A-1 23679 ATTGAAGGGTGGGCGTPACCAGGTTAC 23680GAGTTAATGGGGCTATGCTATTGGCTATTC AC P13069A-1 24588GTCACTATATGGAGTCAAGGTAATTATTGTG TTCAC 24590GGTCTAGAGTTTGAATATTAGTAATGACTTG TATTGAC 24591GCATATTGTGCCATAGAGAGAGAAAATGTAG TAAG P13070A-1 24508CGCTACTGCAAGTTATCAGTCAAGAGATTAT TCC 24510 GCCGTGTAAGCGTGTTTACCAATCTAGTTGP13071A-1 24795 CAAAACTGAGCGAAACTTGTGTTGGGAGAA AG 25395GTGGGGACTCTTTATTCGAAGTTTGCTGAAC P13072A-1 24499CTCATGTAACCAACTCTCTATGAAGTTTGAG ATCCA 24500CTCTAATCGGATTTGGTGTTTCACTTCGGTA AG P13073A-1 24503GATGGCTGTCATTGCTACAGAGGAGTATC 24505 GTGACTCCAAAGGAAAGAGAAATGTTTCTTAAATCATC P13074A-1 25287 GGATAGCAAGTCAATTTCATGCCTTGTGAT AG 25288GCAGGACATGAAGATGTACTTAGTGAATGT GAAG P13158A-1 24713ACTGGAAGAGGGTGCTTAGGGAATCTG 24715 GAGAATCTAGTCTACCACCATACCACGAACP13560A-1 25671 GGGACTCTCTTTATATTGGAAGGTATAACTC AGTG 25672GTTGGACCCTTGTAATTAGACCCGAAACAAA TG P13561A-1 26537ATCTGTCCAACGATCTCTCCATGTTCATTC 27121 CGAATAAGAAGTTGGGTATCACTTACACGTT GGP2447B-2 8815 GATGGGGTTCTAGACTGGGATCTGGAT 8816CTTTTCCTACAGGATTGTCAGGCTTATCGT CA P2481A-1 10468GAAGAGTAACAGAGTCTACGCACCGAC 10470 GTCAACGAACATACTATGCATGATGATTTCT GATTAGP2636C-2 21190 CATATGCAGACAGCAGGCTAAGGAACTC 15824GCAGCAATATAACCAGGATTCAGAATTAATC TAGTTAG P3050A-2 16940CGATGGGGTTGACTTAGAAATGGCATATAC 16860 GCACCAATTCCTCAAGCATACTCCAAACP3436A-1 10660 CTGACAAGGTGTTTTGGTAGGGAGAGATTC 10663GCATCCTCCGTTACTCCAATCAGAGTTTCC AT P3436A-7 10660CTGACAAGGTGTTTTGGTAGGGACACATTC 10663 GCATCCTCCGTTACTCCAATCAGAGTTTCC ATP5219A-1 11913 CACACTATCAACACCTATTGGTGACCATTG TA 8395GGAGGGTGCTTATGTAAATGATGTAAAGAC CAT P5219A-2 11917GATGACCATTTTGATTCCCTCATGCTATTAG TACC 8396CCAATAAGTTAGCAGCATGTGGATCACAGTG TA P5467A-1 19156GTCTCTCGGAGTTGCTTCAATTGCTCATAC 19864 GAGAGTGTGGAATTGTA(AG)TCATTGATTGAAAACTC P5467A-2 19156 GTCTCTCGGAGTTGCTTCAATTGCTCATAC 19864GAGAGTGTGGAATTGTA(AG)TCATTGATTG AAAACTC P6181A-2 17279GCAGAAGGAGCATTGAGGCTTTCCAG 9372 GAAAAGGTTTGTTATGCTTCGTACTCTGTC TCP7659A-1 7817 CATGAAGCTCCACCATTTGCTAGTACATGAA AC 10878CCAGAGTTACCAAACCATCTGTGAGAAATAT CC P7659A-2 7847CATGAAGCTCCACCATTTGCTAGTACATGAA AC 10878 CCAGAGTTACCAAACCATCTGTGAGAAATATCC P8230A-1 13958 CAAACGCTCCCAACAGCTTCAGAATCTC 13959TTGAAGGTTGTAAGAGTCTCGTCGTCG P8524A-1 15081CCCATTCTTCATGTACTCATACACCAAGAG 15086 GCAGCCACCAATAATTTCTCATTTGACAACA AGP8584A-2 19660 CTCTGTATATGAYATATGTGCTCAGTGCCTC 19389GACTTACCAAATGAGTTTGACCAGGTTTTA CC P9026A-1 14494AGGATTCAACCTCTAGCCATGATGATGTTG 14493 CCAAGCTCTTTCCGTGTGTATCAATCTG

APPENDIX IV ASH marker: probes 2 pages Probe 1 Probe 2 Probe 3Marker Name Allele1 sequence Allele2 sequence Allele3 sequence P10355B-118119 GTTTCAGATAAC 18118 GTTTCTGATAAC P10598A-1 13358 GAATGACTTTGA 13360GAATGATTTTGAC P10615A-1 1630 TCATTCTTTCATG 1629 TCATTCATTCATG P10618A-113392 TCTCCAGAAACA 13394 GTTTCGGGAG P10620A-1 13919 GTGATCCGTG 11141ATCACGAATCAC P10621B-2 11150 ATCTTTTCAGGTT 12304 TATGGAGTAATTG P10623A-111360 GGATTACATACTA 11361 TGGATTTTATACTA P10624A-1 13361 TTTGAAGCTTTAT13362 TTTGAATCTTTATC P10632A-1 10697 AAGAATCTTCCTA 10698 AAAGAATATTCCTAP10633A-1 15132 AATCTAAAATTTAGT 15131 ATCTAACATTTAGT P10634A-1 11987ACTAAATTTATACC 11988 GGTATACATTTAG P10635A-1 12562 ATTAGGGGCAG 12563ATTAGGGGGCA P10636A-1 13678 ACTATAGTTCGC 13679 ACTATACTTCGC P10637A-113428 AACCTTTCTGTC 13430 ACCTTGCTGTC P10638B-2 12041 TTGAGGATTTAG 12042TTGAGCATTTAG P10639A-1 15144 TCTCAACTTGGA 15145 TCTCAATTTGGAA P10640A-115137 TTCAGTCAAACC 20442 TTCAGTTAAACC P10641A-1 13673 TTCTTTTGTGACA13675 TTCTTTGGTGAC P10646A-1 6223 TGTGACAACCGA 6224 GTGACACAACCP10648A-1 14935 AATCTTTTTTAAAG 13666 AATCTTCTTTAAAG P10649C-3 11356TCATCTGTGATAA 11358 TCATGTGTGATAA 11357 TCATCTCTGATAA P10651A-1 8002AAGAGAAGGCTA 8003 AAGAGATGGCTA P10782A-1 15302 ATATAAGTAAGGG 15301AATATAAATAAGGG P10783A-1 15599 GCATGTCGAC 15600 GCATGTCGAC P10792A-116759 TTGGAAGTTATAC 15707 TTGGAAGATATAC 15706 TTGGAATATATACT P10793A-115624 GGCATGTGAGT 15625 GGGTTGCGAG P11070A-1 14956 ATTAACAGTAAAGT 14957TATTAACATTAAAGT P11347A-1 15073 TATCTATGTATATTA 17277 TATCTATATATATTAAP12105A-1 23530 TGGGAATGATG 21919 ATCATCCCCAA P12198A-1 23164TAAATAAATAAGATG 23165 AAATAAGTAAGATG P12390B-1 21475 AAAAAAAATGAGG 23524AAAAAATATGAGG P12391A-1 23527 TCAATGTTGGAT 21219 TCAATGATGGATA P12322A-121493 TTGTGACCAATAT 23437 ATATTGATCACAA P12394A-1 23401 ATCAAGCCCAA23528 TGGGTTTGATC P12396A-1 23681 AGAAGCTCGTG 24579 GAAGGTCGTG P13069A-124584 GAAAAAGAAAGG 24585 GAAAAAAAAAGGA 29603 AGATGTTAGAGTTA P13070A-124509 ACATATAATAGTAG 25336 ACATATAGTAGTA P13071A-1 24796 TTTTGTATCTGTAT24798 TTTGTGGCTGTA P13072A-1 25337 TTAACTTGCCAG 25338 TTAACTAGCCAGP13073A-1 24504 AATGATAATTTAGT 24506 AATGATCATTTAG P13074A-1 25306GAATGAATTTTTC 25307 AATGAACTTTTTC P13158A-1 25339 ACACTGCTTAC 25700TTTTTGCTAGAG P13560A-1 25673 ACAACTAATAAGG 25674 TACAACTAAGGTA P13561A-126309 TTCTGATAAAAAAA 26310 TCTGATGAAAAAA P2447B-2 11878 TGTAATGCGTG 8378TGTAACGCGTG 12003 TGTAACGCATGT P2481A-1 5486 GACAATCTAAAAA 7441GACAATTTAAAAA P2636C-2 14670 TAATAATAATTGTGT 15823 AATAATGATTGTGP3050A-2 16760 TTCCTTCTTTTTTT 16761 TCCTTCCTTTTTT P3436A-1 10239GGAACGTTACC 10240 TGGAACATTACC P3436A-7 18078 AATTTTTTAGTATG 14278AATTTTTGAGTATG 14617 ATTTTTTGGTATG P5219A-1 7700 TTATAGACACTTG 7699TATAGGCACTTG P5219A-2 7944 ATAAACCATATATG 7943 ATAAACCTTATATG P5467A-118980 TTAATTACCTTAAG 18981 TTAATTATCTTAAG P5467A-2 20239 ATGCCATTTTG19197 ATGCCCTTTTGT P6181A-2 9448 TAATATCTTATGCA 9292 AATATCGTATGCAP7659A-1 7858 AGTGCGATGAAA 7859 AGTGCACTGAAA P7659A-2 10390 GAGGAGATGTAG7845 AGAGGAGATGTA P8230A-1 10586 TAAAATTGTTGGTT 14933 TAAAATTATTGGTTP8584A-1 14786 TGAAGAAAAATATG 14787 GAACAAAAAGATG P8584A-2 20517CTGTCCACTAA 20358 ACTGTCAACTAA P9026A-1 14340 AGAGGCAGTGA 14341AGAGCCAGTGA

1-40. (canceled)
 41. A plant comprising in its genome a set of geneticelements related to increased yield, wherein said set of geneticelements comprises one or more genetic elements proximal to each of atleast 10% of the markers selected from the group consisting of: Satt684,Satt165, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155,Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480, Satt632-TB,Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409, Satt228,Satt429, Satt426, Satt509, SAT_(—)261, Satt197, Satt519, Satt597, SCT026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1, Satt359, P10648A-1,P12105A-1, P10641A-1, Satt168, Satt556, Satt272, Satt020, Satt066,Satt534, P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB, Satt190,SAT_(—)311-DB, Satt338, Satt227, Satt640-TB, Satt422, Satt457, Satt557,Satt319, SAT_(—)142-DB, Satt460, P13073A-1, Satt307, SCT_(—)028,Satt433, Satt357, Satt321, Satt267, Satt383, Satt295, Satt203, Satt507,SAT_(—)110, P10620A-1, Satt129, Satt147, Satt216, SAT_(—)351, P10621B-2,Satt701, Satt634, Satt558, Satt266, Satt282, Satt537, Satt506, Satt546,P13072A-1, Satt582, Satt389, Satt461, Satt311, Satt514, Satt464,Satt662, Satt543, Satt186, Satt413, Satt672, P13074A-1, P10624A-1,Satt573, Satt598, Satt204, Satt263, Satt491, Satt602, Satt151, Satt355,Satt452, SAT_(—)273-DB, Satt146, Satt193, Satt569, Satt343, Satt586,Satt040, Satt423, Satt348, Satt595, P10782A-1, P3436A-1, P10598A-1,Satt334, Satt510, Satt144, Satt522, P9026A-1, P10646A-1, P5219A-1,P7659A-2, Satt570, Satt356, Satt130, Satt115, Satt594, Satt533, Satt303,Satt352, Satt566, Satt199, Satt503, Satt517, Satt191, SAT_(—)117,Satt353, Satt442, Satt279, Satt314, Satt142, Satt181, Satt367, Satt127,SCTT012, Satt270, Satt292, Satt440, P10640A-1, Satt249, SCT 065,Satt596, Satt280, Satt406, Satt380, Satt183, Satt529, Satt431, Satt242,Satt102, Satt441, Satt544, Satt617, Satt240, P10618A-1, Satt475,Satt196, SAT_(—)301, Satt523, Satt418, Satt398, Satt497, Satt284,Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt590, Satt567,Satt220, Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551,Satt250, Satt346, Satt336, SAT_(—)330-DB, P13069A-1, P5467A-1, P5467A-2,Satt584, SAT_(—)084, P3050A-2, SAT_(—)275-DB, Satt387, Satt549, Satt660,Satt339, Satt255, Satt257, Satt358, P12396A-1, Satt487, Satt259,Satt347, Satt420, Satt576, Satt550, Satt633, Satt262, Satt473, Satt477,Satt581, P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1,P10793A-1, P12391A-1, P13560A-1, P13561A-1, P2481A-1, Satt040, Satt108,Satt109, Satt111, Satt176, Satt219, Satt299, and Satt512.
 42. The plantof claim 41, wherein said set of genetic elements comprises one or moregenetic elements proximal to each of at least 50% of the markersselected from the group consisting of: Satt684, Satt165, Satt042,Satt364, Satt454, Satt526, Satt300, Satt591, Satt155, Satt385, Satt225,Satt236, Satt511, P12390B-1, Satt480, Satt632-TB, Satt233, Satt327,Satt329, Satt508, P10635A-1, Satt409, Satt228, Satt429, Satt426,Satt509, SAT_(—)261, Satt197, Satt519, Satt597, SCT 026, Satt415,Satt583, Satt430, P12198A-1, P8584A-1, Satt359, P10648A-1, P12105A-1,P10641A-1, Satt168, Satt556, Satt272, Satt020, Satt066, Satt534,P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB, Satt190,SAT_(—)311-DB, Satt338, Satt227, Satt640-TB, Satt422, Satt457, Satt557,Satt319, SAT_(—)142-DB, Satt460, P13073A-1, Satt307, SCT_(—)028,Satt433, Satt357, Satt321, Satt267, Satt383, Satt295, Satt203, Satt507,SAT_(—)110, P10620A-1, Satt129, Satt147, Satt216, SAT_(—)351, P10621B-2,Satt701, Satt634, Satt558, Satt266, Satt282, Satt537, Satt506, Satt546,P13072A-1, Satt582, Satt389, Satt461, Satt311, Satt514, Satt464,Satt662, Satt543, Satt186, Satt413, Satt672, P13074A-1, P10624A-1,Satt573, Satt598, Satt204, Satt263, Satt491, Satt602, Satt151, Satt355,Satt452, SAT_(—)273-DB, Satt146, Satt193, Satt569, Satt343, Satt586,Satt040, Satt423, Satt348, Satt595, P10782A-1, P3436A-1, P10598A-1,Satt334, Satt510, Satt144, Satt522, P9026A-1, P10646A-1, P5219A-1,P7659A-2, Satt570, Satt356, Satt130, Satt115, Satt594, Satt533, Satt303,Satt352, Satt566, Satt199, Satt503, Satt517, Satt191, SAT_(—)117,Satt353, Satt442, Satt279, Satt314, Satt142, Satt181, Satt367, Satt127,SCTT012, Satt270, Satt292, Satt440, P10640A-1, Satt249, SCT_(—)065,Satt596, Satt280, Satt406, Satt380, Satt183, Satt529, Satt431, Satt242,Satt102, Satt441, Satt544, Satt617, Satt240, P10618A-1, Satt475,Satt196, SAT_(—)301, Satt523, Satt418, Satt398, Satt497, Satt284,Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt590, Satt567,Satt220, Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551,Satt250, Satt346, Satt336, SAT_(—)330-DB, P13069A-1, P5467A-1, P5467A-2,Satt584, SAT_(—)084, P3050A-2, SAT_(—)275-DB, Satt387, Satt549, Satt660,Satt339, Satt255, Satt257, Satt358, P12396A-1, Satt487, Satt259,Satt347, Satt420, Satt576, Satt550, Satt633, Satt262, Satt473, Satt477,Satt581, P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1,P10793A-1, P12391A-1, P13560A-1, P13561A-1, P2481A-1, Satt040, Satt108,Satt109, Satt111, Satt176, Satt219, Satt299, and Satt512.
 43. The plantof claim 41, wherein said set of genetic elements comprises one or moregenetic elements proximal to each of at least 10% of the markersselected from the group consisting of: Satt642, Satt042, Satt364,Satt454, Satt526, Satt300, Satt591, Satt155, Satt385, Satt511,P12390B-1, Satt632-TB, Satt429, SAT_(—)261, Satt197, P10641A-1, Satt556,Satt534, P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB, Satt190,SAT_(—)311-DB, Satt338, Satt640-TB, Satt557, Satt319, SAT_(—)142-DB,Satt460, Satt433, Satt357, Satt321, Satt295, Satt203, Satt507, Satt129,Satt147, SAT_(—)351, P10621B-2, Satt558, Satt701, Satt634, Satt582,Satt389, Satt464, Satt662, Satt672, Satt573, Satt598, Satt263, Satt602,Satt151, SAT_(—)273-DB, Satt146, Satt193, Satt569, Satt176, Satt343,Satt586, Satt040, Satt595, P10782A-1, Satt334, Satt144, Satt522,Satt570, Satt356, Satt533, Satt199, Satt517, Satt191, SAT_(—)117,Satt279, Satt181, Satt127, Satt270, Satt292, SAT_(—)065, Satt596,Satt406, Satt380, Satt183, Satt529, Satt242, Satt617, Satt240,SAT_(—)301, Satt418, Satt398, Satt497, Satt166, Satt448, Satt373,Satt513, P12394A-1, Satt536, Satt175, Satt677, Satt680, P10615A-1,Satt551, Satt346, Satt336, SAT_(—)330-DB, P13069A-1, P5467A-1, P5467A-2,SAT_(—)084, SAT_(—)275-DB, Satt660, Satt339, P12396A-1, Satt358,Satt487, Satt259, Satt420, Satt576, Satt633, Satt477, Satt581, Satt153,Satt243, P10793A-1, P12391A-1, P12392A-1, P13560A-1, P13561A-1, Satt040,Satt111, Satt176, Satt219, and Satt299.
 44. The plant of claim 41,wherein said set of genetic elements comprises one or more geneticelements proximal to each of at least 10% of the markers selected fromthe group consisting of: Satt684, Satt042, Satt364, Satt454, Satt526,Satt300, Satt591, Satt155, Satt385, Satt632-TB, Satt429, SAT_(—)261,P10641A-1, Satt556, P10638B-2, Satt399, Satt361, Satt661-TB, Satt190,SAT_(—)311-DB, Satt338, Satt640-TB, Satt557, Satt319, SAT_(—)142-DB,Satt321, Satt203, Satt129, Satt147, SAT_(—)351, P10621B-2, Satt701,Satt634, Satt582, Satt389, Satt464, Satt662, Satt672, Satt573, Satt598,Satt263, Satt151, SAT_(—)273-DB, Satt146, Satt193, Satt569, Satt343,Satt586, Satt040, Satt595, Satt334, Satt144, Satt522, Satt570, Satt356,Satt199, Satt517, Satt191, Sat_(—)117, Satt279, Satt181, Satt127,Satt270, Satt292, Sat_(—)065, Satt596, Satt406, Satt380, Satt183,Satt529, Satt242, Satt617, Satt240, SAT_(—)301, Satt418, Satt398,Satt497, Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt536,Satt175, Satt677, Satt680, P10615A-1, Satt551, SAT_(—)330-DB, P13069A-1,P5467A-1, P5467A-2, SAT_(—)084, SAT_(—)275-DB, Satt660, Satt339,Satt358, Satt487, Satt420, Satt576, Satt633, Satt581, Satt153, Satt243,P10793A-1, P13560A-1, P13561A-1, Satt111, Satt219, and Satt299.
 45. Theplant of claim 41, wherein said set of genetic elements comprises one ormore genetic elements proximal to each of at least 50% of the markersselected from the group consisting of: Satt684, Satt526, Satt591,Satt385, Satt632-TB, Satt429, SAT_(—)261, P10641A-1, Satt556, P10638B-2,Satt190, SAT_(—)311-DB, Satt338, Satt640-TB, Satt557, SAT_(—)142-DB,Satt321, Satt203, Satt129, SAT_(—)351, Satt701, Satt582, Satt389,Satt464, Satt672, Satt598, Satt343, Satt595, Satt334, Satt144, Satt522,Satt570, Satt356, Satt199, Sat_(—)117, Satt279, Satt181, Satt127,Satt270, Satt292, Sat_(—)065, Satt529, Satt242, Satt617, SAT_(—)301,Satt398, Satt497, Satt166, Satt373, Satt680, P10615A-1, SAT_(—)330-DB,P13069A-1, SAT_(—)275-DB, Satt339, Satt487, Satt420, Satt58,1 andSatt153.
 46. The plant of claim 41, wherein said set of genetic elementscomprises one or more genetic elements proximal to each of the markers:Satt526, Satt591, Satt429, SAT-_(—)261, P10641A-1, Satt190, Satt557,SAT_(—)142-DB, Satt129, SAT_(—)351, Satt464, Satt343, Satt595, Satt570,Satt181, Satt127, Satt270, Sat_(—)065, Satt529, Satt242, Satt398,Satt166, Satt373, Satt680, SAT_(—)330-DB, P13069A-1, Satt339, Satt487,Satt581, and Satt153.
 47. The plant of claim 41, wherein the plant is asoybean line or soybean variety.
 48. The plant of claim 46, wherein theplant is a soybean line or soybean variety.
 49. The plant of claim 41,wherein the plant comprises an elite soybean strain or an exotic soybeanstrain.
 50. The plant of claim 46, wherein the plant comprises an elitesoybean strain or an exotic soybean strain.
 51. A field comprising aplurality of plants of claim
 41. 52. A field comprising a plurality ofplants of claim
 46. 53. A seed of the plant of claim
 41. 54. A seed ofthe plant of claim
 46. 55. A method of producing a plant havingincreased yield, the method comprising planting the seed of claim 53 insoil and growing a plant therefrom.
 56. A method of producing a planthaving increased yield, the method comprising planting the seed of claim54 in soil and growing a plant therefrom.